Nitrosated and nitrosylated compounds and compositions and their use for treating respiratory disorders

ABSTRACT

Disclosed are (i) compounds of a steroid, a β-agonist, an anticholinergic, a mast cell stabilizer and a phosphodiesterase (PDE) inhibitor directly or indirectly linked to a NO or NO 2  group or a group which stimulates endogenous production of NO or EDRF in vivo; (ii) compositions of steroids, β-agonists, anticholinergics, mast cell stabilizers and PDE inhibitors, which can optionally be substituted with at least one NO or NO 2  moiety or a group which stimulates endogenous production of NO or EDRF in vivo, and a compound that donates, transfers or releases nitric oxide as a charged species, i.e., nitrosonium (NO + ) or nitroxyl (NO − ), or as the neutral species, nitric oxide (NO.) or that stimulates endogenous production of NO or EDRF in vivo; and (iii) uses for them in preventing and/or treating respiratory disorders.

RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No.10/428,936, filed May 5, 2003, which is a divisional of U.S. applicationSer. No. 09/689,851, filed Oct. 13, 2000, issued as U.S. Pat. No.6,579,863, which is a divisional of U.S. application Ser. No.09/157,242, filed Sep. 18, 1998, issued as U.S. Pat. No. 6,197,762,which is a divisional of U.S. application Ser. No. 08/620,882 filed Mar.22, 1996, issued as U.S. Pat. No. 5,824,669, and RE 037611; and is acontinuation of PCT Application No. PCT/US 97/04319 filed Mar. 19, 1997.

BACKGROUND OF THE INVENTION

The present invention relates to the field of compounds, compositionsand uses therefore, in oral and/or nasal administration prophylaxisand/or treatment of respiratory disorders. More particularly theinvention relates to nitrosated and nitrosylated compounds, compositionscomprising such compounds, which can optionally be unsubstituted orsubstituted with at least one NO or NO₂ moiety, and a compound thatdonates, transfers or releases nitric oxide as a charged species, i.e.,nitrosonium (NO+) or nitroxyl (NO), or as the neutral species, nitricoxide (NO.); and uses for each of them.

A broad spectrum of respiratory diseases and disorders have beenrecognized, many of which have overlapping and interacting etiologies.One of the most widespread and prevalent of these diseases in westernpopulations is the chronic disease referred to as “asthma”. Other suchdisorders are also characterized by acute pulmonary vasoconstrictionsuch as may result from pneumonia, traumatic injury, aspiration orinhalation injury, fat embolism in the lung, acidosis inflammation ofthe lung, adult respiratory distress syndrome, acute pulmonary edema,acute mountain sickness, post-cardiac surgery, acute pulmonaryhypertension, persistent pulmonary hypertension of the newborn, prenatalaspiration syndrome, hyaline membrane disease, acute pulmonarythromboembolism, heparin-protamine reactions, sepsis, status asthmaticusor hypoxia (including iatrogenic hypoxia) and other forms of reversiblepulmonary vasoconstriction. Such pulmonary disorders also are alsocharacterized by inflammation of the lung including those associatedwith the migration into the lung of nonresident cell types including thevarious leucocyte subclasses. Also included in the respiratory disorderscontemplated are cystic fibrosis and other diseases which arecharacterized by excess mucosal secretion. Other physiological eventswhich are contemplated to be controlled include platelet activation inthe lung.

Asthma is a major and disabling obstructive respiratory diseaseassociated with significant morbidity and mortality. The term “asthma”has been used to describe a condition which is characterized bywidespread fluctuations in the diameter or caliber of bronchial airwaysover short periods of time resulting in changes in lung function. Theresulting increased resistance to air flow produces symptoms includingbreathlessness (dyspnea), chest constriction or “tightness” and wheeze.The term as used is not currently limited to a disorder or diseaseresulting from any specific cause or causes, rather it is characterizedby its clinical manifestation. A true immunological mechanism may or maynot be a factor in the etiology of an individual asthmatic condition.Further, characteristic wheezing may not be present in particularlysevere attacks where transport of air is completely obstructed.Regardless of the cause, asthma in all sufferers is characterized byreversible hyperresponsiveness of tracheal bronchial smooth muscleresulting in its contraction and interference with normal respiration.The lungs of patients who die of asthma are usually pale pink,hyperinflated, and fail to collapse after their removal from the chest.Many of the airways throughout the bronchial tree are occluded by thickmucus plugs which are infiltrated with various types of leukocytes,including mast cells. The smooth muscle of the airways is hypertrophied.The bronchoconstriction or bronchospasm characterized by asthmaticattacks causes obstruction to air flow which necessitates a forcedexhalation and maintenance of artificially elevated functional airreserve capacity to keep the airways open. The resultant lunghyperinflation places a significant stress on the cardiovascular system(particularly the right ventricle) which can lead to a consequentcardiovascular event. One possible result is a progressive decrease incardiac output referred to as “cardiopulmonary tamponade”. Most deathsresulting from asthma are caused by a condition referred to as “statusasthmaticus,” which is essentially an intensely severe and bronchospasmthat is unresponsive to treatment.

Various categories of drugs are known to be useful in the inhalation oftreatment of asthma. These include β₂ agonists (such as salmeterol,albuterol, metaproternol, terbutaline, pirbuterol, rimiterol,clenbuterol, bitolterol and repreterol, adrenalin, isoproterenol,ephedrine, orciprenlaine, fenoterol and isoetharine); anticholinergicagents (such as atropine, ipratropium, flutropium, tiotropium andrispenzepine) and mast cell stabilizers (chromolyn and nedocromil).Selective β agonists have recently been developed with fewer cardiotonicside effects than those previously employed and are now consideredsuitable therapeutics for management of bronchitis and, particularly,emphysema, for which there previously had not been a suitable effectiveform of therapy.

Although corticosteroids are not generally indicated for routine use inthe treatment of asthma, whether acute or chronic, they are used inlarge doses in the treatment of status asthmaticus. Nonetheless, the useof inhaled corticosteroids for the treatment of bronchial asthma hasincreased in recent years. Most frequently beclomethasone dipropionate,triamcinolone acetonide or flunisolide can be used to reduce or replaceoral corticosteroid therapy, particularly in the treatment of children.This avoids or reduces bronchial reactivity and behavioral toxicity. SeeCott and Cherniack, Steroids and “Steroid-sparing Agents in Asthma”, NewEngl. J. Med., 318:634-636, 1988.

Cystic fibrosis is a multi-organ disorder of the exocrine glands whichis congenital, lethal and affects all populations, particularly Europeanand North American populations. Primary effects of cystic fibrosis arein the secretory glands, particularly mucous secretion. One of the organsystems most effected by cystic fibrosis is the lungs and respiratorytract. Therapy is as yet only symptomatic as the underlying geneticdefect has yet to be characterized.

SUMMARY OF THE INVENTION

The present invention is based on the discovery by the inventors that itis possible to directly or indirectly link an NO or NO₂ group or a groupwhich stimulates the endogenous production of NO or endothelium-derivedrelaxing factor (EDRF) in vivo, to a steroid, a β-agonist, ananticholinergic, a mast cell stabilizer or a phosphodiesterase (PDE)inhibitor and that the resulting compound has beneficial therapeuticeffects of both a steroid, a β-agonist, an anticholinergic, a mast cellstabilizer, or PDE inhibitor and an NO donor or stimulator.

Therefore, one aspect of the invention provides a compound comprising asteroid, a β-agonist, an anticholinergic, a mast cell stabilizer or aPDE inhibitor to which is directly or indirectly linked at least one NOor NO₂ group or a group which stimulates the endogenous production of NOor EDRF in vivo. The groups can be linked through sites such as oxygen(hydroxyl condensation), sulfur (sulfhydryl condensation), carbon andnitrogen.

The invention is further based on the discovery by the inventors that itis possible to co-administer a steroid, a β-agonist, an anticholinergic,a mast cell stabilizer or a PDE inhibitor with a compound that donates,transfers or releases nitric oxide and/or a compound that stimulatesendogenous production of NO or EDRF in vivo. A nitric oxide donor is acompound that contains a nitric oxide moiety and which releases orchemically transfers nitric oxide to another molecule. Nitric oxidedonors include but are not limited to S-nitrosothiols, nitrites,2-hydroxy-2-nitrosohydrazines, and substrates of various forms of nitricoxide synthase. Compounds that stimulate endogenous production of nitricoxide or EDRF in vivo include L-arginine, the substrate for nitric oxidesynthase, cytokines, adenosine, bradykinin, calreticulin, bisacodyl,phenolphthalein, and endothelein.

Therefore, another aspect of the invention provides a compositioncomprising (i) a therapeutically effective amount of a a steroid, aβ-agonist, an anticholinergic, a mast cell stabilizer and/or a PDEinhibitor in combination with (ii) a compound that donates, transfers orreleases nitric oxide and/or a compound that stimulates endogenousproduction of NO or EDRF in vivo.

In another aspect the invention provides a composition comprising (i) atherapeutically effective amount of a steroid, a β-agonist, ananticholinergic, a mast cell stabilizer or a PDE inhibitor to which isdirectly or indirectly linked at least one NO or NO₂ group or a groupthat stimulates endogenous production of NO or EDRF in vivo, and (ii) acompound that donates, transfers or releases nitric oxide and/or acompound that stimulates endogenous production of NO or EDRF in vivo.The invention also provides such compositions in a pharmaceuticallyacceptable carrier.

In another aspect the invention provides a method for treatingrespiratory disorders, such as asthma, in an individual in need thereofwhich comprises administering to the individual a therapeuticallyeffective amount of a steroid, a β-agonist, an anticholinergic, a mastcell stabilizer or a PDE inhibitor to which is directly or indirectlylinked at least one NO or NO₂ group and/or a group that stimulatesendogenous production of NO or EDRF in vivo.

In another aspect the invention provides a method of treatingrespiratory disorders, such as asthma, in an individual in need thereofwhich comprises administering to the individual (i) a therapeuticallyeffective amount of a steroid, a β-agonist, an anticholinergic, a mastcell stabilizer or a PDE inhibitor, which optionally may be substitutedwith at least one NO or NO₂ group or a group that stimulates endogenousproduction of NO or EDRF in vivo, and (ii) a compound that donates,transfers or releases nitric oxide, and/or a group that stimulatesproduction of NO or EDRF in vivo.

The steroid, β-agonist, anticholinergic, mast cell stabilizer or PDEinhibitor and the compound that donates, transfers or releases nitricoxide and/or stimulates endogenous production of NO or EDRF in vivo canbe administered separately or as components of the same composition inone or more pharmaceutically acceptable carriers.

BRIEF DESCRIPTION OF THE FIGURES

The following drawings are illustrative of embodiments of the inventionand do not limit the scope of the invention as defined by the claims.

FIG. 1 illustrates a synthetic pathway for the preparation of nitritecontaining steroid derivatives.

FIG. 2 illustrates a synthetic pathway for the preparation ofnitrosothiol containing steroid derivatives.

FIG. 3 illustrates a synthetic pathway for the preparation of nitratecontaining steroid derivatives.

FIG. 4 illustrates a synthetic pathway for the preparation of2-hydroxy-2-nitrosohydrazine containing steroid derivatives.

FIG. 5 illustrates a synthetic pathway for the preparation of nitritecontaining β-agonist derivatives.

FIG. 6 illustrates a synthetic pathway for the preparation ofnitrosothiol containing β-agonist derivatives.

FIG. 7 illustrates a synthetic pathway for the preparation of nitratecontaining β-agonist derivatives.

FIG. 8 illustrates a synthetic pathway for the preparation of2-hydroxy-2-nitrosohydrazine containing β-agonist derivatives.

FIG. 9 illustrates a synthetic pathway for the preparation of nitritecontaining anticholinergic derivatives.

FIG. 10 illustrates a synthetic pathway for the preparation ofnitrosothiol containing anticholinergic derivatives.

FIG. 11 illustrates a synthetic pathway for the preparation of nitratecontaining anticholinergic derivatives.

FIG. 12 illustrates a synthetic pathway for the preparation of2-hydroxy-2-nitrosohydrazine containing anticholinergic derivatives.

FIG. 13 illustrates a synthetic pathway for the preparation of nitritecontaining rispenzepine derivatives.

FIG. 14 illustrates a synthetic pathway for the preparation ofnitrosothiol containing rispenzepine derivatives.

FIG. 15 illustrates a synthetic pathway for the preparation of nitratecontaining rispenzepine derivatives.

FIG. 16 illustrates a synthetic pathway for the preparation of2-hydroxy-2-nitrosohydrazine containing rispenzepine derivatives.

FIG. 17 illustrates a synthetic pathway for the preparation of nitritecontaining rispenzepine derivatives.

FIG. 18 illustrates a synthetic pathway for the preparation ofnitrosothiol containing rispenzepine derivatives.

FIG. 19 illustrates a synthetic pathway for the preparation of nitratecontaining rispenzepine derivatives.

FIG. 20 illustrates a synthetic pathway for the preparation of2-hydroxy-2-nitrosohydrazine containing rispenzepine derivatives.

FIG. 21 illustrates a synthetic pathway for the preparation of nitritecontaining mast cell stabilizer derivatives.

FIG. 22 illustrates a synthetic pathway for the preparation ofnitrosothiol containing mast cell stabilizer derivatives.

FIG. 23 illustrates a synthetic pathway for the preparation of nitratecontaining mast cell stabilizer derivatives.

FIG. 24 illustrates a synthetic pathway for the preparation of2-hydroxy-2-nitrosohydrazine containing mast cell stabilizerderivatives.

FIG. 25 illustrates a synthetic pathway for the preparation ofsidnonimine containing mast cell stabilizer derivatives.

FIG. 26 illustrates a synthetic pathway for the preparation of nitritecontaining phosphodiesterase inhibitor derivatives.

FIG. 27 illustrates a synthetic pathway for the preparation ofnitrosothiol containing phosphodiesterase inhibitor derivatives.

FIG. 28 illustrates a synthetic pathway for the preparation of nitratecontaining phosphodiesterase inhibitor derivatives.

FIG. 29 illustrates a synthetic pathway for the preparation of2-hydroxy-2-nitrosohydrazine containing phosphodiesterase inhibitorderivatives.

DETAILED DESCRIPTION OF THE INVENTION

The following illustrative elucidations are provided to facilitateunderstanding of certain terms used frequently herein, particularly inthe examples. The elucidations are provided as a convenience and are notlimitative of the invention.

The term “lower alkyl” as used herein refers to branched or straightchain alkyl groups comprising one to ten carbon atoms, including methyl,ethyl, propyl, isopropyl, n-butyl, t-butyl, neopentyl and the like.

The term “alkoxy” as used herein refers to R₅₀O— wherein R₅₀ is loweralkyl as defined above. Representative examples of alkoxy groups includemethoxy, ethoxy, t-butoxy and the like.

The term “alkoxyalkyl” as used herein refers to an alkoxy group aspreviously defined appended to an alkyl group as previously defined.Examples of alkoxyalkyl include, but are not limited to, methoxymethyl,methoxyethyl, isopropoxymethyl and the like.

The term “hydroxy” as used herein refers to —OH.

The term “hydroxyalkyl” as used herein refers to a hydroxy group aspreviously defined appended to a lower alkyl group as previouslydefined.

The term “alkenyl” as used herein refers to a branched or straight chainC₂-C₁₀ hydrocarbon which also comprises one or more carbon-carbon doublebonds.

The term “amino” as used herein refers to —NH₂.

The term “carboxy” as used herein refers to —C(O)O—

The term “nitrate” as used herein refers to —O—NO₂.

The term “alkylamino” as used herein refers to R₅₁NH— wherein R₅₁ is alower alkyl group as defined above, for example, methylamino,ethylamino, butylamino, and the like.

The term “dialkylamino” as used herein refers to R₅₂R₅₃N— wherein R₅₂and R₅₃ are independently selected from lower alkyl groups as definedabove, for example dimethylamino, diethylamino, methyl propylamino andthe like.

The term “nitro” as used herein refers to the group —NO₂ and“nitrosated” refers to compounds that have been substituted therewith.

The term “nitroso” as used herein refers to the group —NO and“nitrosylated” refers to compounds that have been substituted therewith.

The term “aryl” as used herein refers to a mono- or bicyclic carbocyclicring system having one or two aromatic rings including, but not limitedto, phenyl, naphthyl, tetrahydronaphthyl, indanyl, indenyl, and thelike. Aryl groups (including bicyclic aryl groups) can be unsubstitutedor substituted with one, two or three substituents independentlyselected from loweralkyl, haloalkyl, alkoxy, amino, alkylamino,dialkylamino, hydroxy, halo, and nitro. In addition, substituted arylgroups include tetrafluorophenyl and pentafluorophenyl.

The term “arylalkyl” as used herein refers to a lower alkyl radical towhich is appended an aryl group. Representative arylalkyl groups includebenzyl, phenylethyl, hydroxybenzyl, fluorobenzyl, fluorophenylethyl andthe like.

The term “arylalkoxy” as used herein refers to an alkoxy radical towhich is appended an aryl group. Representative arylalkoxy groupsinclude benzyloxy, phenylethoxy, chlorophenylethoxy and the like.

The term “cycloalkyl” as used herein refers to an alicyclic groupcomprising from 3 to 7 carbon atoms including, but not limited to,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and the like.

The term “bridged cycloalkyl” herein refers to two or more cycloalkylradicals fused via adjacent or non-adjacent carbon atoms, including butnot limited to adamantyl and decahydronaphthyl.

The term “cycloalkoxy” as used herein refers to R₅₄O— wherein R₅₄ iscycloalkyl as defined above. Representative examples of alkoxy groupsinclude cyclopropoxy, cyclopentyloxy, and cyclohexyloxy and the like.

The term “arylthio” herein refers to R₅₅S— wherein R₅₅ is an aryl group.

The term “alkylsulfinyl” herein refers to R₅₅—S(O)₂— wherein R₅₅ is aspreviously defined.

The term “carboxamido” herein refers to —C(O)NH₂.

The term “carbamoyl” herein refers to —O—C(O)NH₂.

The term “carboxyl” herein refers to —CO₂H.

The term “halogen” or “halo” as used herein refers to I, Br, Cl, or F.The term “haloalkyl” as used herein refers to a lower alkyl radical, asdefined above, bearing at least one halogen substituent, for example,chloromethyl, fluoroethyl or trifluoromethyl and the like.

The term “haloalkyl” as used herein refers to a lower alkyl radical towhich is appended one or more halogens. Representative examples of ahaloalkyl group include trifluoromethyl, chloromethyl, 2-bromobutyl,1-bromo-2-chloro-pentyl and the like.

The term “haloalkoxy” as used herein refers to a haloalkyl radical towhich is appended an alkoxy group. Representative examples of haloalkoxygroups include fluoromethoxy, 1,1,1-trichloroethoxy, 2-bromobutoxy andthe like.

The term “heteroaryl” as used herein refers to a mono- or bi-cyclic ringsystem containing one or two aromatic rings and containing at least onenitrogen, oxygen, or sulfur atom in an aromatic ring. Heteroaryl groups(including bicyclic heteroaryl groups) can be unsubstituted orsubstituted with one, two or three substituents independently selectedfrom lower alkyl, haloalkyl, alkoxy, amino, alkylamino, dialkylamino,hydroxy, halo and nitro. Examples of heteroaryl groups include but arenot limited to pyridine, pyrazine, pyrimidine, pyridazine, pyrazole,triazole, thiazole, isothiazole, benzothiazole, benzoxazole,thiadiazole, oxazole, pyrrole, imidazole and isoxazole.

The term “heterocyclic ring” refers to any 3-, 4-, 5-, 6-, or 7-memberednonaromatic ring containing at least one nitrogen atom, oxygen, orsulfur.

The term “arylheterocyclic ring” as used herein refers to a bi- ortricyclic ring comprised of an aryl ring as previously defined appendedvia two adjacent carbons of the aryl group to a heterocyclic ring aspreviously defined.

The term “heterocyclic compounds” herein refers to mono and polycycliccompounds containing at least one heteroaryl or heterocyclic ring.

Examples of contemplated steroids include beclamethasone, fluticasone,flunisolide, triamcinolone, butixocort, dexamethasone, fluocortin,budesonide, tixocortal, tipredane and mometasone. Examples ofcontemplated β-agonists include salmeterol, albuterol, metaproterenol,terbutaline, pitbuterol, rimiterol, clenbuterol, bitoterol andreproterol. Examples of contemplated anticholinergics includeipratropium, flutropium, tiotropium and rispenzepine. Examples ofcontemplated mast cell stabilizers include cromalyn and nedocromil.Examples of contemplated PDE inhibitors include filaminast, denbufyllenepiclamilast, zardaverine, and rolipram.

Sources of information for the above include Goodman and Gilman, ThePharmacological Basis of Therapeutics (8th Ed.), McGraw-Hill, Inc.,1993; the Physician's Desk Reference (49th Ed.), Medical Economics(1995); Drug Facts and Comparisons (1993 Ed), Facts and Comparisons(1993); and The Merck Index (11th Ed.), Merck & Co., Inc. (1989), all ofwhich are incorporated herein by reference in their entirety.

A principal aspect of the invention relates to novel nitrosated and/ornitrosylated compounds.

One embodiment of this aspect provides compounds having the structure:

wherein

A is selected from —CH═CH— or —CH₂—CH₂—;

R₁ is selected from

-   -   (1)-C(O)CH₂—B-D wherein B is oxygen or sulfur; D is selected        from (i) —NO; (ii) —NO₂; (iii)        —C(R_(d))—O—C(O)—Y—(C(R_(e))(R_(f)))_(p)-T-Q in which R_(d) is        hydrogen, lower alkyl, cycloalkyl, aryl, alkylaryl, aryl or        heteroaryl, Y is oxygen, sulfur, or NR_(i) in which R_(i) is        hydrogen, lower alkyl, lower haloalkyl, or heteroaryl, R_(e) and        R_(f) are independently selected from hydrogen, lower alkyl,        cycloalkyl, aryl, heteroaryl, arylalkyl, alkylamino,        dialkylamino, carboxy, or taken together are carbonyl,        cycloalkyl or bridged cycloalkyl, p is an integer from 1 to 6, T        is a covalent bond, oxygen, sulfur or nitrogen and Q is selected        from —NO or —NO₂; (iv) —C(O)-T¹-(C(R_(e))(R_(f)))_(p)— T²-Q        wherein T¹ and T² are independently selected from T and R_(e),        R_(f), p, Q, and T are as defined above; (v)        —C(O)-T(C(R_(y))(R_(z)))_(p) wherein R_(y) and R_(e) are        independently selected from        -T¹-(C(R_(e))(R_(f)))_(p)-G-(C(R_(e))(R_(f)))_(p)-T²-Q wherein G        is (i) a covalent bond; (ii) -T-C(O)—; (iii) —C(O)-T, or (iv) Y,        and wherein R_(d), R_(e), R_(f), p, Q, T, and Y are as defined        above;    -   (2) —C(O)—C(O)—O—R_(i) wherein R_(i) is as defined above;    -   (3)-C(O)—B—R_(i) wherein B and R_(i) are as defined above;    -   (4)-C(O)—CH₂—B—C(O)—R_(i) wherein B and R_(i) are as defined        above;    -   (5)-C(O)—CH₂—X wherein X is halogen;    -   (6) —S—R_(i) wherein R_(i) is as defined above;    -   (7)-C(O)CH₂—B-M wherein M is selected from        —C(O)T-(C(R_(e))(R_(f)))_(p)-G-(C(R_(e))(R_(f)))_(p)—N(N—(O⁻)N═O)—R_(i)        or        —C(R_(d))—O—C(O)T-(C(R_(e))(R_(f)))_(p)-G-(C(R_(e))(R_(f)))_(p)—N(N—(O—)N═O)—R_(i)        wherein R_(e), R_(f), R₁, p, G and T are as defined above;

R₂ and R₃ are independently selected from hydrogen, hydroxyl, loweralkyl, —O(O)C—R_(i), or —S—R_(i) wherein R_(i) is as defined above or R₂and R₃ when taken together are

wherein R¹ _(i) and R² _(i) are independently selected from R_(i)wherein R_(i) is as defined above;

R₄ and R₅ are independently selected from hydrogen or halogen;

-   R₆ is selected from hydrogen, D, or M wherein D and M are as defined    above with the provision that R₆ must be D or M if the selection for    R₁ does not include D or M.

Another embodiment of this aspect provides compounds having thestructure:

wherein,

E is nitrogen or C—R₇ wherein R₇ is hydrogen, halogen, —CH₂O—R_(j), or—O—R_(j) wherein R_(j) is hydrogen, D or M wherein D and M are definedas above;

R₈ and R₉ are independently selected from amino, hydrogen, —CH₂O—R_(j),or —O—R_(k) wherein R_(k) is —C(O)—R_(d) or R_(j) and R_(d) and R_(j)are as defined above;

S is (1)-CH₂—N(Z)-R₁₀ wherein Z is hydrogen, —(N(O—)N═O), or M wherein Mis as defined above and R₁₀ is selected from

-   -   (i) lower alkyl    -   (ii) —(CH₂)_(p)—O—(CH₂)_(a)—C₆H₅ wherein a is an integer from 1        to 4 and p is as defined above;    -   wherein Z is as defined above; and

R₁₁ is selected from hydrogen, D, or M with the provision that R₁₁ mustbe D or M if neither R₈ or R₉ include D or M and Z is hydrogen.

Another embodiment of this aspect provides compounds having thestructure:K is a a monovalent charged anion selected from halide, nitrate, ornitrite;

wherein,

K is a a monovalent charged anion selected from halide, nitrate, ornitrite;

R₁₂ and R₁₃ are hydrogen or, when taken together are oxygen;

R₁₄ is lower alkyl or haloalkyl;

R₁₅ is selected from:

wherein R_(v) is selected from D or M and D and M are as defined above.

Another embodiment of this aspect provides compounds having thestructure:

wherein

R₁₄ is as defined above;

R₁₆ is selected from hydrogen,—C(R_(d))—O—C(O)—Y—(C(R_(e))(R_(f)))_(p)-T-Q,—C(O)-T¹—(C(R_(e))(R_(f)))_(p)— T²-Q, or M; and wherein R_(d), R_(e),R_(f), M, p, T, T¹, T², Q, and Y are defined as above; and

R₁₇ and R₁₈ are independently selected from a lone pair of electrons,—C(R_(d))—O—C(O)—Y—(C(R_(e))(R_(f)))_(p)-T-Q, or M wherein R_(d), R_(e),R_(f), M, p, T, Q, and Y are defined as above with the provision thatR₁₇ and/or R₁₈ must be —C(R_(d))—O—C(O)—Y—(C(R_(e))(R_(f)))_(p)-T-Q or Mwhen R₁₆ is hydrogen.

Another embodiment of this aspect provides compounds having thestructure:

wherein R₁₉ is selected from—(C(R_(e))(R_(f)))_(p)-G-(C(R_(e))(R_(f)))_(p)-T-Q or—S(O₂)—(C(R_(e))(R_(f)))_(p)-G-(C(R_(e))(R_(f)))_(p)—N(N—(O—)N═O)—R_(i)wherein R_(e), R_(f), R_(i), p, G, Q and T are as defined above; andwherein R₁₄, R₁₇, and R₁₈ are as defined above.

Another embodiment of this aspect provides compounds having thestructure:

wherein,

F is selected from oxygen or NR_(i) wherein R_(i) is as defined above;

R₂₀ and R₂₁ are independently selected from (1)—Y—(C(R_(e))(R_(f)))_(p)—H—(C(R_(e))(R_(f)))_(p)-T-Q; wherein H is (i) acovalent bond; (ii) -T-C(O)—; (iii) —C(O)-T; (iv)—C(Y—C(O)—R_(m))_(p)—wherein R_(m) is heteroaryl or heterocyclic ring;and wherein Y, R_(d), R_(e), R_(f), p, Q and T are as defined above; (2)T-(C(R_(e))(R_(f)))_(p)—H—(C(R_(e))(R_(f)))_(p)—N(N—(O—)N═O)—R_(i)wherein R_(d), R_(e), R_(f), R_(i), p, H and T are as defined above; (3)

in which W is a heterocyclic ring or NR_(s)R′_(s) wherein R_(s) andR′_(s) are independently selected from lower alkyl, aryl or alkenyl; (4)sodium or (5) hydrogen;

-   R₂₂ is hydrogen, M, or D with the provision that R₂₂ must be M or D    when R₂₀ and R₂₁ are selected as sodium or hydrogen.

Another embodiment of this aspect provides compounds having thestructure:

wherein,

R₂₃ is alkoxy, cycloalkoxy, or halogen;

R₂₄ is hydrogen, alkoxy, or haloalkoxy; and

R₂₅ is selected from:

wherein R₂₆ is selected from D or M and wherein R₁₆, R₁₈, and R₁₉ aredefined as above.

Compounds of the invention which have one or more asymmetric carbonatoms may exist as the optically pure enantiomers, pure diastereomers,mixtures of enantiomers, mixtures of diastereomers, racemic mixtures ofenantiomers, diastereomeric racemates or mixtures of diastereomericracemates. It is to be understood that the present invention anticipatesand includes within its scope all such isomers and mixtures thereof.

Another aspect of the invention provides processes for making the novelcompounds of the invention and to the intermediates useful in suchprocesses.

Some of the compounds of the invention are synthesized as shown in FIGS.1 through 29 presented below, in which A, E, F, W, R₂, R₃, R₄, R₅, R₆,R₉, R₁₀, R₁₂, R₁₃, R₁₄, R₁₇, R₁₈, R₂₁, R₂₂, R₂₃, R₂₄, R_(e), R_(f), p,and Z are as defined above or as depicted in the reaction schemes forstructures I-VII; P¹ is an oxygen protecting group, P² is a sulfurprotecting group and P³ is a nitrogen protecting group. The reactionsare performed in solvents appropriate to the reagents and materialsemployed are suitable for the transformations being effected. It isunderstood by those skilled in the art of organic synthesis that thefunctionality present in the molecule must be consistent with thechemical transformation proposed. This will, on occasion, necessitatejudgment by the routine as to the order of synthetic steps, protectinggroups required, and deprotection conditions. Substituents on thestarting materials may be incompatible with some of the reactionconditions required in some of the methods described, but alternativemethods and substituents compatible with the reaction conditions will bereadily apparent to skilled practitioners in the art. The use of sulfur,oxygen, and nitrogen protecting groups is well known in the art forprotecting thiol, alcohol, and amino groups against undesirablereactions during a synthetic procedure and many such protecting groupsare known, c.f., T. H. Greene and P. G. M. Wuts, Protective Groups inOrganic Synthesis, John Wiley & Sons, New York (1991).

One embodiment of this aspect provides processes for making compoundshaving structures I and to the intermediates useful in such processes asfollows.

Nitroso compounds of formula (I) wherein A, R₂, R₃, R₄, R₅, R₆, R_(e),R_(f), and p are defined as above and an O-nitrosylated ester isrepresentative of the R₁ group as defined above may be prepared asoutlined in FIG. 1. The alcohol group of formula 1 is converted to theester of formula 2 wherein p, R_(e) and R_(f) are defined as above byreaction with an appropriate protected alcohol containing activatedacylating agent wherein P¹ is as defined above. Preferred methods forthe formation of esters are reacting the alcohol with the preformed acidchloride or symmetrical anhydride of the protected alcohol containingacid or condensing the alcohol and protected alcohol containing acid inthe presence of a dehydrating agent such as dicyclohexylcarbodiimide(DCC) or 1-ethyl-3(3-dimethylaminopropyl)carbodiimide hydrochloride(EDAC HCl) with or without a catalyst such as dimethylaminopyridine(DMAP) or 1-hydroxybenzotriazole (HOBt). Preferred protecting groups forthe alcohol moiety are silyl ethers such as a trimethylsilyl or atert-butyldimethylsilyl ether. Deprotection of the hydroxyl moiety(fluoride ion is the preferred method for removing silyl etherprotecting groups) followed by reaction a suitable nitrosylating agentsuch as thionyl chloride nitrite, thionyl dinitrate, or nitrosoniumtetrafluoroborate in a suitable anhydrous solvent such asdichloromethane, THF, DMF, or acetonitrile with or without an amine basesuch as pyridine or triethylamine affords the compound of the formulaIA.

Nitroso compounds of formula (I) wherein A, R₂, R₃, R₄, R₅, R₆, R_(e),R_(f), and p are defined as above and an S-nitrosylated ester isrepresentative of the R₁ group as defined above may be prepared asoutlined in FIG. 2. The alcohol group of the formula 1 is converted tothe ester of the formula 3 wherein p, R_(e) and R_(f) are defined asabove by reaction with an appropriate protected thiol containingactivated acylating agent wherein p² is as defined above. Preferredmethods for the formation of esters are reacting the alcohol with thepreformed acid chloride or symmetrical anhydride of the protected thiolcontaining acid or condensing the alcohol and protected thiol containingacid in the presence of a dehydrating agent such as DCC or EDAC HCl withor without a catalyst such as DMAP or HOBt. Preferred protecting groupsfor the thiol moiety are as a thioester such as a thioacetate orthiobenzoate, as a disulfide, as a thiocarbamate such as N-methoxymethylthiocarbamate, or as a thioether such as a paramethoxybenzyl thioether,a tetrahydropyranyl thioether or a 2,4,6-trimethoxybenzyl thioether.Deprotection of the thiol moiety (zinc in dilute aqueous acid,triphenylphosphine in water and sodium borohydride are preferred methodsfor reducing disulfide groups while aqueous base is typically utilizedto hydrolyze thioesters and N-methoxymethyl thiocarbamates and mercurictrifluoroacetate, silver nitrate, or strong acids such astrifluoroacetic or hydrochloric acid and heat are used to remove aparamethoxybenzyl thioether, a tetrahydropyranyl thioether, or a2,4,6-trimethoxybenzyl thioether group) followed by reaction a suitablenitrosylating agent such as thionyl chloride nitrite, thionyl dinitrate,a lower alkyl nitrite such as tert-butyl nitrite, or nitrosoniumtetrafluoroborate in a suitable anhydrous solvent such as methylenechloride, THF, DMF, or acetonitrile with or without an amine base suchas pyridine or triethylamine affords the compound of the formula IB.Alternatively, treatment of compound 3 with a stoichiometric quantity ofsodium nitrite in an acidic aqueous or alcoholic solution affords thecompound of the formula IB.

Nitro compounds of formula (I) wherein A, R₂, R₃, R₄, R₅, R₆, R_(e),R_(f), and p are defined as above and an O-nitrosated ester isrepresentative of the R₁ group as defined above may be prepared asoutlined in FIG. 3. The alcohol group of the formula 1 is converted tothe ester of the formula IC wherein p, R_(e) and R_(f) are defined asabove by reaction with an appropriate nitrate containing activatedacylating agent. Preferred methods for the formation of esters arereacting the alcohol with the preformed acid chloride or symmetricalanhydride of the nitrate containing acid or condensing the alcohol andnitrate containing acid in the presence of a dehydrating agent such asDCC or EDAC HCl with or without a catalyst such as DMAP or HOBt toafford a compound of the formula IC.

2-Hydroxy-2-nitrosohydrazine compounds of formula (I) wherein A, R₂, R₃,R₄, R₅, R₆, R_(e), R_(f), R_(i) and p are defined as above and a2-hydroxy-2-nitrosohydrazine ester is representative of the R₁ group asdefined above may be prepared as outlined in FIG. 4. The alcohol groupof the formula 1 is converted to the ester of the formula 4 wherein p,R_(e), R_(f), and R₁ are defined as above by reaction with anappropriate protected amino containing activated acylating agent whereinP³ is as defined above. Preferred methods for the formation of estersare reacting the alcohol with the preformed acid chloride or symmetricalanhydride of the protected amino containing acid or condensing thealcohol and protected amine containing acid in the presence of adehydrating agent such as DCC or EDAC. HCl with or without a catalystsuch as DMAP or HOBt. Preferred protecting groups for the amine are as acarbamate such as a t-butyl carbamate or a 9-fluorenylmethyl carbamateor an amide such as a trifluoroacetamide. Deprotection of the aminomoiety (strong acid such as HCl in dioxane or trifluoroacetic acid isused to remove a t-butyl carbamate while piperidine is used to remove9-fluorenylmethyl carbamate while mild aqueous or alcoholic base may beused to cleave a trifluoroacetamide group) followed by treatment of theamine with nitric oxide (1-5 atmospheres) in a dry inert solvent such asether or tetrahydrofuran affords the compound of the formula ID.

Another embodiment of this aspect provides processes for makingcompounds having structures II and to the intermediates useful in suchprocesses as follows.

Nitroso compounds of formula (II) wherein E, R₉, R₁₀, R_(e), R_(f), andp are defined as above, Z and R₁₁ are hydrogen, and an O-nitrosylatedester is representative of the R₈ group as defined above may be preparedas outlined in FIG. 5. The amine, 1⁰, and 2⁰ alcohol groups of formula 5are protected to afford the compound of the formula 6. Preferredprotecting groups for the amine are as a carbamate such as a benzylcarbamate or an amide such as a trifluoroacetamide while preferredprotecting groups for 1⁰ and 2⁰ alcohol moieties are as benzyl ethers.The phenolic group(s) of formula 6 is converted to the ester(s) offormula 7 wherein p, R_(e) and R_(f) are defined as above by reactionwith an appropriate protected alcohol containing activated acylatingagent wherein P¹ is as defined above. Preferred methods for theformation of esters are reacting the alcohol with the preformed acidchloride or symmetrical anhydride of the protected alcohol containingacid or condensing the alcohol and protected alcohol containing acid inthe presence of a dehydrating agent such as DCC or EDAC HCl with orwithout a catalyst such as DMAP or HOBt. Preferred protecting groups forthe alcohol moiety are silyl ethers such as a trimethylsilyl ortert-butyldimethylsilyl ether. Deprotection of the hydroxyl moieties(fluoride ion is the preferred method for removing silyl etherprotecting groups) followed by reaction a suitable nitrosylating agentsuch as thionyl chloride nitrite, thionyl dinitrate, or nitrosoniumtetrafluoroborate in a suitable anhydrous solvent such asdichloromethane, THF, DMF, or acetonitrile with or without an amine basesuch as pyridine or triethylamine affords the compound of the formula 8.The compound of the formula 8 is then converted to the compound of theformula IIA by deprotecting the amine and remaining hydroxyl groups.Hydrogen in the presence of a transition metal catalyst such aspalladium or platinum is a preferred method for removing benzyl etherand benzyl carbamate protecting groups.

Nitroso compounds of formula (II) wherein E, R₉, R₁₀, R_(e), R_(f), andp are defined as above, Z and R₁₁ are hydrogen, and a S-nitrosylatedester is representative of the R₈ group as defined above may be preparedas outlined in FIG. 6. The amine, 1⁰, and 2⁰ alcohol groups of formula 5are protected to afford the compound of the formula 9. Preferredprotecting groups for the amine are as a carbamate such as a t-butylcarbamate or an amide such as a trifluoroacetamide while preferredprotecting groups for 1⁰ and 2⁰ alcohol moieties are as silyl etherssuch as trimethylsilyl or t-butyldimethylsilyl ethers. The phenolicgroup(s) of the formula 9 is converted to the ester(s) of the formula 10wherein p, R_(e) and R_(f) are defined as above by reaction with anappropriate protected thiol containing activated acylating agent whereinP² is as defined above. Preferred methods for the formation of estersare reacting the alcohol with the preformed acid chloride or symmetricalanhydride of the protected thiol containing acid or condensing thealcohol and protected thiol containing acid in the presence of adehydrating agent such as DCC or EDAC HCl with or without a catalystsuch as DMAP or HOBt. Preferred protecting groups for the thiol moietyare as a thioester such as a thioacetate or thiobenzoate, as adisulfide, as a thiocarbamate such as N-methoxymethyl thiocarbamate, oras a thioether such as a paramethoxybenzyl thioether, atetrahydropyranyl thioether or a 2,4,6-trimethoxybenzyl thioether.Deprotection of the amine (strong acid such as HCl in dioxane ortrifluoroacetic acid is used to remove a t-butyl carbamate while mildaqueous or alcoholic base is used to cleave trifluoroacetamide groups)and hydroxyl moieties (fluoride ion is the preferred method for removingsilyl ether protecting groups) followed by deprotection of the thiolmoiety (zinc in dilute aqueous acid, triphenylphosphine in water andsodium borohydride are preferred methods for reducing disulfide groupswhile aqueous base is typically utilized to hydrolyze thioesters andN-methoxymethyl thiocarbamates and mercuric trifluoroacetate, silvernitrate, or strong acids such as trifluoroacetic or hydrochloric acidand heat are used to remove a paramethoxybenzyl thioether, atetrahydropyranyl thioether or a 2,4,6-trimethoxybenzyl thioether group)affords a compound of the formula 11. Reaction of the compound of theformula 11 with a an equimolar equivalent based upon thiol of a suitablenitrosylating agent such as thionyl chloride nitrite, thionyl dinitrate,a lower alkyl nitrite such as tert-butyl nitrite, or nitrosoniumtetrafluoroborate in a suitable anhydrous solvent such as methylenechloride, THF, DMF, or acetonitrile with or without an amine base suchas pyridine or triethylamine affords the compound of the formula IIB.Alternatively, treatment of compound 11 with a stoichiometric quantityof sodium nitrite in an acidic aqueous or alcoholic solution affords thecompound of the formula IIB.

Nitro compounds of formula (II) wherein E, R₉, R₁₀, R_(e), R_(f), and pare defined as above, Z and R₁₁ are hydrogen, and a O-nitrosated esteris representative of the R₈ group as defined above may be prepared asoutlined in FIG. 7. The amine, 1⁰, and 2⁰ alcohol groups of formula 5are protected to afford the compound of the formula 12. Preferredprotecting groups for the amine are as a carbamate such as a t-butylcarbamate or an amide such as a trifluoroacetamide while preferredprotecting groups for 1⁰ and 2⁰ alcohol moieties are as silyl etherssuch as trimethylsilyl or t-butyldimethylsilyl ethers. The phenolicgroup(s) of the formula 12 is converted to the ester(s) of the formula13 wherein p, R_(e) and R_(f) are defined as above by reaction with anappropriate nitrate containing activated acylating agent. Preferredmethods for the formation of esters are reacting the alcohol with thepreformed acid chloride or symmetrical anhydride of the protected thiolcontaining acid or condensing the alcohol and protected thiol containingacid in the presence of a dehydrating agent such as DCC or EDAC HCl withor without a catalyst such as DMAP or HOBt. Deprotection of the amine(strong acid such as HCl in dioxane or trifluoroacetic acid is used toremove a t-butyl carbamate) and hydroxyl moieties (fluoride ion is thepreferred method for removing silyl ether protecting groups) affords acompound of the formula IIC.

2-Hydroxy-2-nitrosohydrazine compounds of formula (II) wherein E, Z, R₉,R₁₀, R_(e), R_(f), R_(i), and p are defined as above and a2-hydroxy-2-nitrosohydrazine ester is representative of the R₈ group asdefined above may be prepared as outlined in FIG. 8. The amine, 1⁰, and2⁰ alcohol groups of formula 5 are protected to afford the compound ofthe formula 12. Preferred protecting groups for the amine are as acarbamate such as a t-butyl carbamate or an amide such as atrifluoroacetamide while preferred protecting groups for 1⁰ and 2⁰alcohol moieties are as silyl ethers such as trimethylsilyl ort-butyldimethylsilyl ethers. The phenolic group(s) of the formula 12 isconverted to the ester of the formula 14 wherein p, R_(e) and R_(f) aredefined as above by reaction with an appropriate protected aminocontaining activated acylating agent wherein P³ is as defined above.Preferred methods for the formation of esters are reacting the alcoholwith the preformed acid chloride or symmetrical anhydride of theprotected amino containing acid or condensing the alcohol and protectedamine containing acid in the presence of a dehydrating agent such as DCCor EDAC. HCl with or without a catalyst such as DMAP or HOBt. Preferredprotecting groups for the amine are as a carbamate such as a t-butylcarbamate or an amide such as a trifluoroacetamide. Deprotection of theamine(s) (strong acid such as HCl in dioxane or trifluoroacetic acid isused to remove a t-butyl carbamate while mild aqueous or alcoholic baseis used to cleave trifluoroacetamide groups) and hydroxyl moieties(fluoride ion is the preferred method for removing silyl etherprotecting groups) followed by treatment of the amine(s) with nitricoxide (1-5 atmospheres) in a dry inert solvent such as ether ortetrahydrofuran affords the compound of the formula IID.

Another embodiment of this aspect provides processes for makingcompounds having structures III and to the intermediates useful in suchprocesses as follows.

Nitroso compounds of formula (III) wherein aryl, heteroaryl, R₁₂, R₁₃,R₁₄, R_(e), R_(f), and p are defined as above and an O-nitrosylatedester is representative of the R₁₅ group as defined above may beprepared as outlined in FIG. 9. The alcohol group of formula 15 isconverted to the ester of formula 16 wherein p, R_(e) and R_(f) aredefined as above by reaction with an appropriate protected alcoholcontaining activated acylating agent wherein P¹ is as defined above.Preferred methods for the formation of esters are reacting the alcoholwith the preformed acid chloride or symmetrical anhydride of theprotected alcohol containing acid or condensing the alcohol andprotected alcohol containing acid in the presence of a dehydrating agentsuch as DCC or EDAC. HCl with or without a catalyst such as DMAP orHOBt. Preferred protecting groups for the alcohol moiety are silylethers such as a tert-butyldimethylsilyl ether. Deprotection of thehydroxyl moiety (fluoride ion is the preferred method for removing silylether protecting groups) followed by reaction a suitable nitrosylatingagent such as thionyl chloride nitrite, thionyl dinitrate, ornitrosonium tetrafluoroborate in a suitable anhydrous solvent such asdichloromethane, THF, DMF, or acetonitrile with or without an amine basesuch as pyridine or triethylamine affords the compound of the formulaIIIA.

Nitroso compounds of formula (III) wherein aryl, heteroaryl, R₁₂, R₁₃,R₁₄, R_(e), R_(f), and p are defined as above and an S-nitrosylatedester is representative of the R₁₅ group as defined above may beprepared as outlined in FIG. 10. The alcohol group of the formula 15 isconverted to the ester of the formula 17 wherein p, R_(e) and R_(f) aredefined as above by reaction with an appropriate protected thiolcontaining activated acylating agent wherein P² is as defined above.Preferred methods for the formation of esters are reacting the alcoholwith the preformed acid chloride or symmetrical anhydride of theprotected thiol containing acid or condensing the alcohol and protectedthiol containing acid in the presence of a dehydrating agent such as DCCor EDAC HCl with or without a catalyst such as DMAP or HOBt. Preferredprotecting groups for the thiol moiety are as a thioester such as athioacetate or thiobenzoate, as a disulfide, as a thiocarbamate such asN-methoxymethyl thiocarbamate, or as a thioether such as aparamethoxybenzyl thioether, a tetrahydropyranyl thioether or a2,4,6-trimethoxybenzyl thioether. Deprotection of the thiol moiety (zincin dilute aqueous acid, triphenylphosphine in water and sodiumborohydride are preferred methods for reducing disulfide groups whileaqueous base is typically utilized to hydrolyze thioesters andN-methoxymethyl thiocarbamates and mercuric trifluoroacetate, silvernitrate, or strong acids such as trifluoroacetic or hydrochloric acidand heat are used to remove a paramethoxybenzyl thioether, atetrahydropyranyl thioether, or a 2,4,6-trimethoxybenzyl thioethergroup) followed by reaction a suitable nitrosylating agent such asthionyl chloride nitrite, thionyl dinitrate, a lower alkyl nitrite suchas tert-butyl nitrite, or nitrosonium tetrafluoroborate in a suitableanhydrous solvent such as methylene chloride, THF, DMF, or acetonitrilewith or without an amine base such as pyridine or triethylamine affordsthe compound of the formula IIIB. Alternatively, treatment of compound17 with a stoichiometric quantity of sodium nitrite in an acidic aqueousor alcoholic solution affords the compound of the formula IIIB.

Nitro compounds defined ester of formula (III) wherein aryl, heteroaryl,R₁₂, R₁₃, R₁₄, R_(e), R_(f), and p are defined as above and anO-nitrosated ester is representative of the R₁₅ group as defined abovemay be prepared as outlined in FIG. 11. The alcohol group of the formula15 is converted to the ester of the formula IIIC wherein p, R_(e) andR_(f) are defined as above by reaction with an appropriate nitratecontaining activated acylating agent. Preferred methods for theformation of esters are reacting the alcohol with the preformed acidchloride or symmetrical anhydride of the nitrate containing acid orcondensing the alcohol and nitrate containing acid in the presence of adehydrating agent such as DCC or EDAC HCl with or without a catalystsuch as DMAP or HOBt to afford a compound of the formula IIIC.

2-Hydroxy-2-nitrosohydrazine compounds of formula (III) wherein aryl,heteroaryl, R₁₂, R₁₃, R₁₄, R_(e), R_(f), R_(i), and p are defined asabove and a 2-hydroxy-2-nitrosohydrazine ester is representative of theR₁₅ group as defined above may be prepared as outlined in FIG. 12. Thealcohol group of the formula 15 is converted to the ester of the formula18 wherein p, R_(e), R_(f), and R_(i) are defined as above by reactionwith an appropriate protected amino containing activated acylating agentwherein P³ is as defined above. Preferred methods for the formation ofesters are reacting the alcohol with the preformed acid chloride orsymmetrical anhydride of the protected amino containing acid orcondensing the alcohol and protected amine containing acid in thepresence of a dehydrating agent such as DCC or EDAC. HCl with or withouta catalyst such as DMAP or HOBt. Preferred protecting groups for theamine are as a carbamate such as a t-butyl carbamate or a9-fluorenylmethyl carbamate or an amide such as a trifluoroacetamide.Deprotection of the amino moiety (strong acid such as HCl in dioxane ortrifluoroacetic acid is used to remove a t-butyl carbamate whilepiperidine is used to remove 9-fluorenylmethyl carbamate while mildaqueous or alcoholic base may be used to cleave a trifluoroacetamidegroup) followed by treatment of the amine with nitric oxide (1-5atmospheres) in a dry inert solvent such as ether or tetrahydrofuranaffords the compound of the formula IIID.

Another embodiment of this aspect provides processes for makingcompounds having structures IV and to the intermediates useful in suchprocesses as follows.

Nitroso compounds of formula (IV) wherein R₁₄, R₁₇, R₁₈, R_(e), R_(f),and p are defined as above and an O-nitrosylated imide is representativeof the R₁₆ group as defined above may be prepared as outlined in FIG.13. The amide nitrogen of formula 19 is converted to the imide offormula 20 wherein p, R_(e) and R_(f) are defined as above by reactionwith an appropriate protected alcohol containing activated acylatingagent wherein P¹ is as defined above. Preferred methods for theformation of imides are reacting the alcohol with the preformed acidchloride or symmetrical anhydride of the protected thiol containing acidor condensing the alcohol and protected thiol containing acid in thepresence of a dehydrating agent such as DCC or EDAC. HCl with a catalystsuch as DMAP. Preferred protecting groups for the alcohol moiety aresilyl ethers such as a tert-butyldimethylsilyl ether. Deprotection ofthe hydroxyl moiety (fluoride ion is the preferred method for removingsilyl ether protecting groups) followed by reaction a suitablenitrosylating agent such as thionyl chloride nitrite, thionyl dinitrate,or nitrosonium tetrafluoroborate in a suitable anhydrous solvent such asdichloromethane, THF, DMF, or acetonitrile with or without an amine basesuch as pyridine or triethylamine affords the compound of the formulaIVA.

Nitroso compounds of formula (IV) wherein R₁₄, R₁₇, R₁₈, R_(e), R_(f),and p are defined as above and an S-nitrosylated imide is representativeof the R₁₆ group as defined above may be prepared as outlined in FIG.14. The amide nitrogen of formula 19 is converted to the imide of theformula 21 wherein p, R_(e) and R_(f) are defined as above by reactionwith an appropriate protected thiol containing activated acylating agentwherein P² is as defined above. Preferred methods for the formation ofimides are reacting the alcohol with the preformed acid chloride orsymmetrical anhydride of the protected thiol containing acid orcondensing the alcohol and protected thiol containing acid in thepresence of a dehydrating agent such as DCC or EDAC HCl with a catalystsuch as DMAP. Preferred protecting groups for the thiol moiety are as athioester such as a thioacetate or thiobenzoate, as a disulfide, or as athioether such as a paramethoxybenzyl thioether, a tetrahydropyranylthioether or a 2,4,6-trimethoxybenzyl thioether. Deprotection of thethiol moiety (zinc in dilute aqueous acid, triphenylphosphine in waterand sodium borohydride are preferred methods for reducing disulfidegroups while mercuric trifluoroacetate, silver nitrate, or strong acidssuch as trifluoroacetic or hydrochloric acid and heat are used to removea paramethoxybenzyl thioether, a tetrahydropyranyl thioether, or a2,4,6-trimethoxybenzyl thioether group) followed by reaction a suitablenitrosylating agent such as thionyl chloride nitrite, thionyl dinitrate,a lower alkyl nitrite such as tert-butyl nitrite, or nitrosoniumtetrafluoroborate in a suitable anhydrous solvent such as methylenechloride, THF, DMF, or acetonitrile with or without an amine base suchas pyridine or triethylamine affords the compound of the formula IVB.Alternatively, treatment of compound 21 with a stoichiometric quantityof sodium nitrite in an acidic aqueous or alcoholic solution affords thecompound of the formula IVB.

Nitro compounds of formula (IV) wherein R₁₄, R₁₇, R₁₈, R_(e), R_(f), andp are defined as above and an O-nitrosated imide is representative ofthe R₁₆ group as defined above may be prepared as outlined in FIG. 15.The amide of the formula 19 is converted to the imide of the formula IVCwherein p, R_(e) and R_(f) are defined as above by reaction with anappropriate nitrate containing activated acylating agent. Preferredmethods for the formation of esters are reacting the alcohol with thepreformed acid chloride or symmetrical anhydride of the nitratecontaining acid or condensing the alcohol and nitrate containing acid inthe presence of a dehydrating agent such as DCC or EDAC HCl with acatalyst such as DMAP to afford a compound of the formula IVC.

2-Hydroxy-2-nitrosohydrazine compounds of formula (IV) wherein R₁₄, R₁₇,R₁₈, R_(e), R_(f), R_(i), and p are defined as above and a2-hydroxy-2-nitrosohydrazine imide is representative of the R₁₆ group asdefined above may be prepared as outlined in FIG. 16. The amide nitrogenof the formula 19 is converted to the imide of the formula 22 wherein p,R_(e), R_(f), and R_(i) are defined as above by reaction with anappropriate protected amino containing activated acylating agent whereinP³ is as defined above. Preferred methods for the formation of imidesare reacting the amide with the preformed acid chloride or symmetricalanhydride of the protected amino containing acid or condensing thealcohol and protected amine containing acid in the presence of adehydrating agent such as DCC or EDAC. HCl with a catalyst such as DMAP.Preferred protecting groups for the amine are as a carbamate such as at-butyl carbamate or a 9-fluorenylmethyl carbamate. Deprotection of theamino moiety (strong acid such as HCl in dioxane or trifluoroacetic acidis used to remove a t-butyl carbamate while piperidine is used to remove9-fluorenylmethyl carbamate) followed by treatment of the amine withnitric oxide (1-5 atmospheres) in a dry inert solvent such as ether ortetrahydrofuran affords the compound of the formula IVD.

Another embodiment of this aspect provides processes for makingcompounds having structures V and to the intermediates useful in suchprocesses as follows.

Nitroso compounds of formula (V) wherein R₁₄, R₁₇, R₁₈, R_(e), R_(f),and p are defined as above and an O-nitrosylated isothioamide isrepresentative of the R₁₉ group as defined above may be prepared asoutlined in FIG. 17. The amide of formula 19 is converted to thethioamide of formula 23 wherein by reaction with a thiation reagent suchas 2,4-bis(4-methoxyphenyl)-2,4-dithioxo-1,2,3,4-dithiadiphosphetane orphosphorus pentasulfide. Alkylation of compound of the formula 23 withan appropriate protected alcohol containing alkylating agent affords acompound of the formula 24 wherein p, R_(e), R_(f), and P¹ are definedas above. Preferred alkylating agents are alkyl halides or sulfonateswhile preferred protecting groups for the alcohol moiety are silylethers such as a trimethylsilyl or a tert-butyldimethylsilyl ether.Deprotection of the hydroxyl moiety (fluoride ion is the preferredmethod for removing silyl ether protecting groups) followed by reactiona suitable nitrosylating agent such as thionyl chloride nitrite, thionyldinitrate, or nitrosonium tetrafluoroborate in a suitable anhydroussolvent such as dichloromethane, THF, DMF, or acetonitrile with orwithout an amine base such as pyridine or triethylamine affords thecompound of the formula VA.

Nitroso compounds of formula (V) wherein R₁₄, R₁₇, R₁₈, R_(e), R_(f),and p are defined as above and an S-nitrosylated isothioamide isrepresentative of the R₁₉ group as defined above may be prepared asoutlined in FIG. 18. Alkylation of compound of the formula 23 with anappropriate protected thiol containing alkylating agent affords acompound of the formula 25 wherein p, R_(e), R_(f), and P² are definedas above. Preferred alkylating agents are alkyl halides or sulfonateswhile preferred protecting groups for the thiol moiety are as athioester such as a thioacetate or thiobenzoate, as a disulfide, or as athioether such as a 2,4,6-trimethoxybenzyl thioether. Deprotection ofthe thiol moiety (zinc in dilute aqueous acid, triphenylphosphine inwater and sodium borohydride are preferred methods for reducingdisulfide groups, mild aqueous or alcoholic base is used to cleavethioesters strong acids such as trifluoroacetic or hydrochloric acid areused to remove a 2,4,6-trimethoxybenzyl thioether group) followed byreaction with a suitable nitrosylating agent such as thionyl chloridenitrite, thionyl dinitrate, a lower alkyl nitrite such as tert-butylnitrite, or nitrosonium tetrafluoroborate in a suitable anhydroussolvent such as methylene chloride, THF, DMF, or acetonitrile with orwithout an amine base such as pyridine or triethylamine affords thecompound of the formula VB. Alternatively, treatment of compound 25 witha stoichiometric quantity of sodium nitrite in an acidic aqueous oralcoholic solution affords the compound of the formula VB.

Nitro compounds of formula (V) wherein R₁₄, R₁₇, R₁₈, R_(e), R_(f), andp are defined as above and an O-nitrosated isothioamide isrepresentative of the R₁₉ group as defined above may be prepared asoutlined in FIG. 19. The thioamide of the formula 23 is converted to thecompound of the formula VC wherein p, R_(e) and R_(f) are defined asabove by reaction with an appropriate nitrate containing alkylatingagent. Preferred alkylating agents are alkyl halides or sulfonates.

2-Hydroxy-2-nitrosohydrazine compounds of formula (V) wherein R₁₄, R₁₇,R₁₈, R_(e), R_(f), R_(i), and p are defined as above and a2-hydroxy-2-nitrosohydrazine isothioamide is representative of the R₁₉group as defined above may be prepared as outlined in FIG. 20. Thethioamide of the formula 23 is converted to the compound of the formula26 by reaction with an appropriate protected amino containing alkylatingagent wherein P³ is as defined above. Preferred alkylating agents arealkyl halides or sulfonates while preferred protecting groups for theamine are as a carbamate such as a t-butyl carbamate or a9-fluorenylmethyl carbamate. Deprotection of the amino moiety (strongacid such as HCl in dioxane or trifluoroacetic acid is used to remove at-butyl carbamate while piperidine is used to remove 9-fluorenylmethylcarbamate) followed by treatment of the amine with nitric oxide (1-5atmospheres) in a dry inert solvent such as ether or tetrahydrofuranaffords the compound of the formula VD.

Another embodiment of this aspect provides processes for makingcompounds having structures VI and to the intermediates useful in suchprocesses as follows.

Nitroso compounds of formula (VI) wherein F, R₂₁, R₂₂, R_(e), R_(f), andp are defined as above and an O-nitrosylated ester is representative ofthe R₂₃ group as defined above may be prepared as outlined in FIG. 21.The alcohol and acid groups of formula 27 are protected to afford thecompound of the formula 28. Preferred protecting groups for the alcoholare as a carbamate such as a benzyl carbamate or a formate ester such asa benzoylformate ester while preferred protecting groups for the acidsare as esters such as t-butyl esters. Deprotection of the hydroxylmoiety (catalytic hydrogenation is the preferred method for cleavingbenzyl carbamates while mild aqueous base removes the benzoylformateester group) followed by reaction of the alcohol group with anappropriate protected alcohol containing activated acylating agentwherein R_(e), R_(f), and p and P¹ is as defined above affords acompound of the formula 29. Preferred methods for the formation ofesters are reacting the alcohol with the preformed acid chloride orsymmetrical anhydride of the protected alcohol containing acid orcondensing the alcohol and protected alcohol containing acid in thepresence of a dehydrating agent such as DCC or EDAC. HCl with or withouta catalyst such as DMAP or HOBt. Preferred protecting groups for thealcohol moiety are silyl ethers such as a tert-butyldimethylsilyl ether.Deprotection of the acid and hydroxyl moieties (strong acid such as HClin dioxane or trifluoroacetic acid cleaves t-butyl esters while fluorideion is the preferred method for removing silyl ether protecting groups)followed by reaction a suitable nitrosylating agent such as thionylchloride nitrite, thionyl dinitrate, or nitrosonium tetrafluoroborate ina suitable anhydrous solvent such as dichloromethane, THF, DMF, oracetonitrile with or without an amine base such as pyridine ortriethylamine affords the compound of the formula VIA.

Nitroso compounds of formula (VI) wherein F, R₂₁, R₂₂, R_(e), R_(f), andp are defined as above and an S-nitrosylated ester is representative ofthe R₂₃ group as defined above may be prepared as outlined in FIG. 22.The compound of the formula 30 is converted to the compound of theformula 31 by reaction of the alcohol group with an appropriateprotected thiol containing activated acylating agent wherein R_(e),R_(f), and p and P² is as defined above. Preferred methods for theformation of esters are reacting the alcohol with the preformed acidchloride or symmetrical anhydride of the protected thiol containing acidor condensing the alcohol and protected thiol containing acid in thepresence of a dehydrating agent such as DCC or EDAC HCl with or withouta catalyst such as DMAP or HOBt. Preferred protecting groups for thethiol moiety are as a thioester such as a thioacetate or thiobenzoate,as a disulfide, as a thiocarbamate such as N-methoxymethylthiocarbamate, or as a thioether such as a paramethoxybenzyl thioether,a tetrahydropyranyl thioether or a 2,4,6-trimethoxybenzyl thioether.Deprotection of the thiol and acid moieties (zinc in dilute aqueousacid, triphenylphosphine in water and sodium borohydride are preferredmethods for reducing disulfide groups while aqueous base is typicallyutilized to hydrolyze thioesters and N-methoxymethyl thiocarbamates andmercuric trifluoroacetate, silver nitrate, or strong acids such astrifluoroacetic or hydrochloric acid and heat are used to remove aparamethoxybenzyl thioether, a tetrahydropyranyl thioether, or a2,4,6-trimethoxybenzyl thioether group as well as t-butyl esters)followed by reaction a suitable nitrosylating agent such as thionylchloride nitrite, thionyl dinitrate, a lower alkyl nitrite such astert-butyl nitrite, or nitrosonium tetrafluoroborate in a suitableanhydrous solvent such as methylene chloride, THF, DMF, or acetonitrilewith or without an amine base such as pyridine or triethylamine affordsthe compound of the formula VIB. Alternatively, treatment of thedeprotected compound with a stoichiometric quantity of sodium nitrite inan acidic aqueous or alcoholic solution affords the compound of theformula VIB.

Nitroso compounds of formula (VI) wherein F, R₂₁, R₂₂, R_(e), R_(f), andp are defined as above and an O-nitrosated ester is representative ofthe R₂₃ group as defined above may be prepared as outlined in FIG. 23.The alcohol group of the formula 30 is converted to the ester of theformula 32 wherein p, R_(e) and R_(f) are defined as above by reactionwith an appropriate nitrate containing activated acylating agent.Preferred methods for the formation of esters are reacting the alcoholwith the preformed acid chloride or symmetrical anhydride of the nitratecontaining acid or condensing the alcohol and nitrate containing acid inthe presence of a dehydrating agent such as DCC or EDAC HCl with orwithout a catalyst such as DMAP or HOBt. Deprotection of the acidmoieties (strong acid such as HCl in dioxane or trifluoroacetic acidcleaves t-butyl esters) affords the compound of the formula VIC.

2-Hydroxy-2-nitrosohydrazine compounds of formula (VI) wherein F, R₂₁,R₂₂, R_(e), R_(f), R_(i), and p are defined as above and a2-hydroxy-2-nitrosohydrazine ester is representative of the R₂₃ group asdefined above may be prepared as outlined in FIG. 24. The alcohol groupof the formula 30 is converted to the ester of the formula 33 wherein p,R_(e) and R_(f) and P³ are defined as above by reaction with anappropriate protected amine containing activated acylating agent.Preferred methods for the formation of esters are reacting the alcoholwith the preformed acid chloride or symmetrical anhydride of theprotected amino containing acid or condensing the alcohol and protectedamine containing acid in the presence of a dehydrating agent such as DCCor EDAC. HCl with or without a catalyst such as DMAP or HOBt. Preferredprotecting groups for the amine are as a carbamate such as a t-butylcarbamate or a 9-fluorenylmethyl carbamate or an amide such ad such as atrifluoroacetamide. Deprotection of the amino and t-butyl ester moieties(strong acid such as HCl in dioxane or trifluoroacetic acid is used toremove a t-butyl carbamate while piperidine is used to remove9-fluorenylmethyl carbamate while mild aqueous or alcoholic base may beused to cleave a trifluoroacetamide group and strong acid such as HCl indioxane or trifluoroacetic acid is used to remove a t-butyl estergroups) followed by treatment of the amine with nitric oxide (1-5atmospheres) in a dry inert solvent such as ether or tetrahydrofuranaffords the compound of the formula VID.

Sidnonimine compounds of formula (VI) wherein F, R₂₂, R_(e), R_(f), andp are defined and a 6-W-substituted sydnonimine wherein W is as definedabove is representative of the R₂₀ and R₂₁ groups as defined above maybe prepared as outlined in FIG. 25. The alcohol of formula 27 isprotected to afford the compound of the formula 34. Preferred protectinggroups for the alcohol are as a carbamate such as a t-butyl carbonate ora silyl ether such as a trimethylsilyl ether. The diacid of the formula34 is converted into the dicarboximide of the formula 35 by reactionwith a 6-W-substituted sydnonimine. Preferred methods for thepreparation of carboximides are initially forming the dimixed anhydridevia reaction of 34 with a chloroformate such as isobutylchloroformate inthe presence of a non nucleophilic base such as triethylamine in ananhydrous inert solvent such as diethylether or THF. The dimixedanhydride is then reacted with the 6-W-substituted sydnonimine to afford35. Alternatively, the diacid 34 and may be coupled to the6-W-substituted sydnonimine afford 35 by treatment with a dehydrationagent such as DCC or EDAC with or without a catalyst such as DMAP orHOBt. Alternatively, the diacid 34 may be converted into an active esterby reaction with a suitably substituted phenol utilizing any of theconditions for ester formation described, followed by reaction with thea 6-W-substituted sydnonimine. Preferred 6-W-substituted sydnoniminesare 1,2,6,4-oxatriazolium, 6-amino-6-morpholine and are1,2,6,4-oxatriazolium, 6-amino-6-(6-chloro-2-methyl-benzene) andpreferred active esters are para-nitrophenyl, 2,4,6-trichlorophenyl, andpentafluorophenyl. Deprotection of the hydroxyl moiety (strong acid suchas HCl or trifluoroacetic acid is used to cleave t-butyl carbonateswhile fluoride is the preferred method for removing silyl ethers) in thecompound of the formula 35 affords the compound of the formula VIE.

Another embodiment of this aspect provides processes for makingcompounds having structures VII and to the intermediates useful in suchprocesses as follows.

Nitroso compounds of formula (VII) wherein R₂₃, R₂₄, R_(e), R_(f), and pare defined as above and an O-nitrosylated imide is representative ofthe R₂₅ group as defined above may be prepared as outlined in FIG. 26.The amide nitrogen of formula 36 is converted to the imide of formula 37wherein p, R_(e) and R_(f) are defined as above by reaction with anappropriate protected alcohol containing activated acylating agentwherein P¹ is as defined above. Preferred methods for the formation ofimides are reacting the alcohol with the preformed acid chloride orsymmetrical anhydride of the protected alcohol containing acid orcondensing the alcohol and protected alcohol containing acid in thepresence of a dehydrating agent such as DCC or EDAC. HCl with a catalystsuch as DMAP. Preferred protecting groups for the alcohol moiety aresilyl ethers such as a tert-butyldimethylsilyl ether. Deprotection ofthe hydroxyl moiety (fluoride ion is the preferred method for removingsilyl ether protecting groups) followed by reaction a suitablenitrosylating agent such as thionyl chloride nitrite, thionyl dinitrate,or nitrosonium tetrafluoroborate in a suitable anhydrous solvent such asdichloromethane, THF, DMF, or acetonitrile with or without an amine basesuch as pyridine or triethylamine affords the compound of the formulaVIIA.

Nitroso compounds of formula (VII) wherein R₂₃, R₂₄, R_(e), R_(f), and pare defined as above and an S-nitrosylated imide is representative ofthe R₂₅ group as defined above may be prepared as outlined in FIG. 27.The amide nitrogen of formula 36 is converted to the imide of formula 38wherein p, R_(e) and R_(f) are defined as above by reaction with anappropriate protected thiol containing activated acylating agent whereinp² is as defined above. Preferred methods for the formation of imidesare reacting the amide with the preformed acid chloride or symmetricalanhydride of the protected thiol containing acid or condensing the amideand protected thiol containing acid in the presence of a dehydratingagent such as DCC or EDAC. HCl with a catalyst such as DMAP. Preferredprotecting groups for the thiol moiety are as a thioester such as athioacetate or thiobenzoate, as a disulfide, as a thiocarbamate such asN-methoxymethyl thiocarbamate, or as a thioether such as aparamethoxybenzyl thioether, a tetrahydropyranyl thioether or a2,4,6-trimethoxybenzyl thioether. Deprotection of the thiol moiety (zincin dilute aqueous acid, triphenylphosphine in water and sodiumborohydride are preferred methods for reducing disulfide groups whileaqueous base is typically utilized to hydrolyze thioesters andN-methoxymethyl thiocarbamates and mercuric trifluoroacetate, silvernitrate, or strong acids such as trifluoroacetic or hydrochloric acidand heat are used to remove a paramethoxybenzyl thioether, atetrahydropyranyl thioether, or a 2,4,6-trimethoxybenzyl thioethergroup) followed by reaction a suitable nitrosylating agent such asthionyl chloride nitrite, thionyl dinitrate, a lower alkyl nitrite suchas tert-butyl nitrite, or nitrosonium tetrafluoroborate in a suitableanhydrous solvent such as methylene chloride, THF, DMF, or acetonitrilewith or without an amine base such as pyridine or triethylamine affordsthe compound of the formula VIIB. Alternatively, treatment of compound38 with a stoichiometric quantity of sodium nitrite in an acidic aqueousor alcoholic solution affords the compound of the formula VIIB.

Nitro compounds of formula (VII) wherein R₂₃, R₂₄, R_(e), R_(f), and pare defined as above and an O-nitrosated imide is representative of theR₂₅ group as defined above may be prepared as outlined in FIG. 28. Theamide of the formula 36 is converted to the imide of the formula VIICwherein p, R_(e) and R_(f) are defined as above by reaction with anappropriate nitrate containing activated acylating agent. Preferredmethods for the formation of esters are reacting the alcohol with thepreformed acid chloride or symmetrical anhydride of the nitratecontaining acid or condensing the alcohol and nitrate containing acid inthe presence of a dehydrating agent such as DCC or EDAC HCl with acatalyst such as DMAP.

2-Hydroxy-2-nitrosohydrazine compounds of formula (VII) wherein R₂₃,R₂₄, R_(e), R_(f), R_(i), and p are defined as above and a2-hydroxy-2-nitrosohydrazine imide is representative of the R₂₅ group asdefined above may be prepared as outlined in FIG. 29. The amide of theformula 36 is converted to the imide of the formula 39 wherein p, R_(e),R_(f), and R_(i) are defined as above by reaction with an appropriateprotected amino containing activated acylating agent wherein P³ is asdefined above. Preferred methods for the formation of imides arereacting the amide with the preformed acid chloride or symmetricalanhydride of the protected amino containing acid or condensing thealcohol and protected amine containing acid in the presence of adehydrating agent such as DCC or EDAC:HCl with a catalyst such as DMAP.Preferred protecting groups for the amine are as a carbamate such as at-butyl carbamate or a 9-fluorenylmethyl carbamate. Deprotection of theamino moiety (strong acid such as HCl in dioxane or trifluoroacetic acidis used to remove a t-butyl carbamate while piperidine is used to remove9-fluorenylmethyl carbamate) followed by treatment of the amine withnitric oxide (1-5 atmospheres) in a dry inert solvent such as ether ortetrahydrofuran affords the compound of the formula VIID.

As noted above, another aspect the invention provides a compositioncomprising (i) a therapeutically effective amount of a steroid, aβ-agonist, an anticholinergic, a mast cell stabilizer or a PDEinhibitor, which optionally can be substituted with at least one NO orNO₂ group or a group that stimulates endogenous production of NO or EDRFin vivo, and (ii) a compound that donates, transfers or releases nitricoxide as a charged species, i.e., nitrosonium (NO⁺) or nitroxyl (NO⁻),or as the neutral species, nitric oxide (NO.) and/or a compound thatstimulates endogenous production of NO or EDRF in vivo.

The compounds that donate, transfer or release nitric oxide can be anyof those known to the art, including those mentioned and/or exemplifiedbelow.

Nitrogen monoxide can exist in three forms: NO⁻ (nitroxyl), NO. (nitricoxide) and NO⁺ (nitrosonium). NO. is a highly reactive short-livedspecies that is potentially toxic to cells. This is critical, becausethe pharmacological efficacy of NO depends upon the form in which it isdelivered. In contrast to NO⁻ nitrosonium and nitroxyl do not react withO₂ ⁻ species, and are also resistant to decomposition in the presence ofredox metals. Consequently, administration of NO equivalents does notresult in the generation of toxic by-products or the elimination of theactive NO moiety.

Compounds contemplated for use in the invention are nitric oxide andcompounds that release nitric oxide or otherwise directly or indirectlydeliver or transfer nitric oxide to a site of its activity, such as on acell membrane, in vivo. As used here, the term “nitric oxide”encompasses uncharged nitric oxide (NO.) and charged nitric oxidespecies, particularly including nitrosonium ion (NO⁺) and nitroxyl ion(NO⁻). The reactive form of nitric oxide can be provided by gaseousnitric oxide. The nitric oxide releasing, delivering or transferringcompounds, having the structure K—NO wherein K is a nitric oxidereleasing, delivering or transferring moiety, include any and all suchcompounds which provide nitric oxide to its intended site of action in aform active for their intended purpose. As used here, the term “NOadducts” encompasses any of such nitric oxide releasing, delivering ortransferring compounds, including, for example, S-nitrosothiols,nitrothiols, O-nitrosoalcohols, O-nitroalcohols, sydnonimines,2-hydroxy-2-nitrosohydrazines (NONOates),(E)-alkyl-2-((E)-hydroxyimino)-5-nitro-3-hexene amines or amides,nitrosoamines, furoxans, as well a substrates for the endogenous enzymeswhich synthesize nitric oxide. It is contemplated that any or all ofthese “NO adducts” can be mono- or poly-nitrosylated or nitrosated at avariety of naturally susceptible or artificially provided binding sitesfor nitric oxide or derivatives which donate or release NO. One group ofsuch NO adducts is the S-nitrosothiols, which are compounds that includeat least one —S—NO group. Such compounds include S-nitroso-polypeptides(the term “polypeptide” includes proteins and also polyamino acids thatdo not possess an ascertained biological function, and derivativesthereof); S-nitrosylated amino acids (including natural and syntheticamino acids and their stereoisomers and racemic mixtures and derivativesthereof); S-nitrosylated sugars, S-nitrosylated-modified and unmodifiedoligonucleotides (preferably of at least 5, and more particularly 5-200,nucleotides); and an S-nitrosylated hydrocarbons where the hydrocarboncan be a branched or unbranched, and saturated or unsaturated aliphatichydrocarbon, or an aromatic hydrocarbon; S-nitrosylated hydrocarbonshaving one or more substituent groups in addition to the S-nitrosogroup; and heterocyclic compounds. S-nitrosothiols and the methods forpreparing them are described in U.S. Pat. No. 5,380,758; Oae et al.,Org. Prep. Proc. Int., 15(3):165-198 (1983); Loscalzo et al., J.Pharmacol. Exp. Ther., 249(3):726729 (1989) and Kowaluk et al., J.Pharmacol. Exp. Ther., 256:1256-1264 (1990), all of which areincorporated in their entirety by reference.

One particularly preferred embodiment of this aspect relates toS-nitroso amino acids where the nitroso group is linked to a sulfurgroup of a sulfur-containing amino acid or derivative thereof. Forexample, such compounds include the following:S-nitroso-N-acetylcysteine, S-nitroso-captopril, S-nitroso-homocysteine,S-nitroso-cysteine and S-nitroso-glutathione.

Suitable S-nitrosylated proteins include thiol-containing proteins(where the NO group is attached to one or more sulfur group on an aminoacid or amino acid derivative thereof) from various functional classesincluding enzymes, such as tissue-type plasminogen activator (TPA) andcathepsin B; transport proteins, such as lipoproteins, heme proteinssuch as hemoglobin and serum albumin; and biologically protectiveproteins, such as the immunoglobulins and the cytokines. Suchnitrosylated proteins are described in PCT Publ. Applic. No. WO93/09806, published May 27, 1993. Examples include polynitrosylatedalbumin where multiple thiol or other nucleophilic centers in theprotein are modified.

Further examples of suitable S-nitrosothiols include those having thestructures:

(i) CH₃(C(R_(e))(R_(f)))_(x)SNO

wherein x equals 2 to 20 and R_(e) and R_(f) are as defined above;

(ii) HS(C((R_(e))(R_(f)))_(x)SNO

wherein x equals 2 to 20; and R_(e) and R_(f) are as defined above;

(iii) ONS(C(R_(e))(R_(f)))_(x)B; and

(iv) H₂N—(CO₂H)—(CH₂)_(x)—C(O)NH—C(CH₂SNO)—C(O)NH—CH₂—CO₂H

wherein x equals 2 to 20; R_(e) and R_(f) are as defined above; and B isselected from the group consisting of fluoro, C₁-C₆ alkoxy, cyano,carboxamido, cycloalkyl, arylalkoxy, alkylsulfinyl, arylthio,alkylamino, dialkylamino, hydroxy, carbamoyl, N-alkylcarbamoyl,N,N-dialkylcarbamoyl, amino, hydroxyl, carboxyl, hydrogen, nitro andaryl.

Nitrosothiols can be prepared by various methods of synthesis. Ingeneral, the thiol precursor is prepared first, then converted to theS-nitrosothiol derivative by nitration of the thiol group with NaNO₂under acidic conditions (pH is about 2.5) to yield the S-nitrosoderivative. Acids which may be used for this purpose include aqueoussulfuric, acetic and hydrochloric acids. Alternatively, the precursorthiol may be nitrosylated by treatment with an alkyl nitrite such astert-butyl nitrite.

Another group of such NO adducts are those wherein the compounds donate,transfer or release nitric oxide and are selected from the groupconsisting of compounds that include at least one ON—N— or ON—C— group.The compound that includes at least one ON—N— or ON—C— group ispreferably selected from the group consisting of ON—N— orON—C-polypeptides (the term “polypeptide” includes proteins and alsopolyamino acids that do not possess an ascertained biological function,and derivatives thereof); ON—N— or ON—C-amino acids (including naturaland synthetic amino acids and their stereoisomers and racemic mixtures);ON—N— or ON—C-sugars; ON—N— or ON—C-modified and unmodifiedoligonucleotides (preferably of at least 5, and more particularly 5-200,nucleotides), ON—O—, ON—N— or ON—C-hydrocarbons which can be branched orunbranched, saturated or unsaturated aliphatic hydrocarbons or aromatichydrocarbons; ON—N— or ON—C— hydrocarbons having one or more substituentgroups in addition to the ON—N— or ON—C— group; and ON—N— orON—C-heterocyclic compounds.

Another group of such NO adducts is the nitrites which have an —O—NOgroup wherein the organic template to which the nitrite group isappended is a protein, polypeptide, amino acid, carbohydrate, branchedor unbranched and saturated or unsaturated alkyl, aryl or a heterocycliccompound. A preferred example is the nitrosylated form of isosorbide.Compounds in this group form S-nitrosothiol intermediates in vivo in therecipient human or other animal to be treated and can therefore includeany structurally analogous precursor R—O—NO of the S-nitrosothiolsdescribed above.

Another group of such adducts are nitrates which donate, transfer orrelease nitric oxide and are selected from the group consisting ofcompounds that include at least one at least one O₂N—O—, O₂N—N—, O₂N—S—or O₂N—C— group. Preferred among these are those selected from the groupconsisting of O₂N—O—, O₂N—N—, O₂N—S— or O₂N—C-polypeptides; O₂N—O—,O₂N—N—, O₂N—S— or O₂N—C-amino acids; O₂N—O—, O₂N—N—O₂N—S— orO₂N—C-sugars; O₂N—O—, O₂N—N—, O₂N—S— or O₂N—C-modified and unmodifiedoligonucleotides; O₂N—O—, O₂N—N—, O₂N—S— or O₂N—C— hydrocarbons whichcan be branched or unbranched, saturated or unsaturated aliphatichydrocarbons or aromatic hydrocarbons; O₂N—O—, O₂N—N—, O₂N—S— or O₂N—C—hydrocarbons having one or more substituent groups in addition to theO₂N—O—, O₂N—N—, O₂N—S— or O₂N—C— group; and O₂N—O—, O₂N—N—, O₂N—S— orO₂N—C-heterocyclic compounds. Preferred examples are isosorbidedinitrate and isosorbide mononitrate.

Another group of such NO adducts is the nitroso-metal compounds whichhave the structure (L)_(u)-M-(NO)_(v)-A_(z). M is a metal, preferably atransition metal; L includes any appropriate organic or inorganic liganddirectly attached to the metal; u, v, z are each integers independentlyselected from Ito 5; and A is an appropriate organic or inorganiccounterion to yield a neutral species. Preferred metals include iron,copper, manganese, cobalt, selenium and luthidium. A preferred exampleis sodium nitroprusside.

Another group of such adducts are 2-hydroxy-2-nitrosohydrazines whichdonate, transfer or release nitric oxide and have a R₆₁R₆₂N—N(O-M⁺)—NOgroup wherein R₆₁ and R₆₂ include polypeptides, amino acids, sugars,modified and unmodified oligonucleotides, hydrocarbons where thehydrocarbon can be a branched or unbranched, and saturated orunsaturated aliphatic hydrocarbon or an aromatic hydrocarbon,hydrocarbons having one or more substituent groups and heterocycliccompounds. M⁺ is a metal cation, such as, for example, a Group I metalcation.

Another group of such adducts are thionitrates which donate, transfer orrelease nitric oxide and have the structure R₆₁—S—NO₂ wherein R₆₁ is asdescribed above.

Compounds that stimulate endogenous synthesis of NO or EDRF in vivoinclude L-arginine, the substrate for nitric oxide synthase, cytokines,adenosine, bradykinin, calreticulin, bisacodyl, phenolphthalein, andendothelein.

The compounds and compositions of the invention are useful in thetreatment of a large number of disorders, particularly those related toor characterized by pulmonary dysfunction. Examples of such indicationsinclude preventing (if given prior to the onset of symptoms) orreversing the following: acute pulmonary vasoconstriction, such as mayresult from pneumonia, traumatic injury, aspiration of inhalationinjury, fat embolism in the lung, acidosis, inflammation of the lung,adult respiratory distress syndrome, acute pulmonary edema, acutemountain sickness, asthma, post cardiac surgery acute pulmonaryhypertension, persistent pulmonary hypertension of the newborn, prenatalaspiration syndrome, hyaline membrane disease, acute pulmonarythromboembolism, heparin-protamine reactions, sepsis, asthma, statusasthmaticus, or hypoxia (including that which may occur during one-lunganesthesia), as well as those cases of chronic pulmonaryvasoconstriction which have a reversible component, such as may resultfrom chronic pulmonary hypertension, bronchopulmonary dysplasia, chronicpulmonary thromboembolism, idiopathic or primary pulmonary hypertension,or chronic hypoxia.

When administered in vivo, the nitric oxide adduct may be administeredin combination with pharmaceutical carriers and in dosages describedherein.

The instant disclosure, particularly in combination with the significantbody of literature, commercially available pharmaceutically acceptableliquid, solid and gaseous carriers and vehicles, volume expanders,tabletting and encapsulation materials, enteric and other coatings, andinhalant and intranasal delivery devices and the ordinary skill of thosepracticing in the field, amply teaches the reader how to use thecompounds and compositions of the invention in the methods of theinvention and particularly for oral and nasal inhalation and intranasaltherapy. The following is supplementary to and exemplary thereof.

The pharmaceutical compositions utilized in this invention can beadministered preferably by inhalation (oral and/or nasal), and also byintranasal mucosal administration, oral, enteral, topical, vaginal,sublingual, rectal, intramuscular, intravenous, or subcutaneous means.

The compounds of this invention can be employed in combination withconventional excipients; i.e., pharmaceutically acceptable organic orinorganic carrier substances suitable for parenteral, enteral orintranasal application which do not deleteriously react with the activecompounds. Suitable pharmaceutically acceptable carriers include, butare not limited to, water, salt solutions, alcohol, vegetable oils,polyethylene glycols, gelatin, lactose, amylose, magnesium stearate,talc, silicic acid, viscous paraffin, perfume oil, fatty acidmonoglycerides and diglycerides, petroethral fatty acid esters,hydroxymethylcellulose, polyvinylpyrrolidone, etc. The pharmaceuticalpreparations can be sterilized and if desired, mixed with auxiliaryagents, e.g., lubricants, preservatives, stabilizers, wetting agents,emulsifiers, salts for influencing osmotic pressure, buffers, colorings,flavoring and/or aromatic substances and the like which do notdeleteriously react with the active compounds.

Many therapeutically active ingredients have been administered orapplied to the body by means of inhalation, such as in aerosol dosageform. One of the main reasons for the rapid and widespread acceptance ofthe inhalation dosage forms for the administration of therapeuticallyactive agents is that it affords many and distinct advantages to theuser. These advantages have been described by various investigators and,for aerosol and metered-dose inhalers, include rapid onset of action;circumvention of the first-pass effect; avoidance of degradation in thegastrointestinal tract; lower dosage which will minimize adversereactions; dose titration to individual needs and ideal for as-neededmedication. It is an ideal alternate route when the chosen therapeuticagent may interact chemically or physically with other medicinals neededconcurrently, or when the drug entity exhibits erratic pharmacokineticsupon oral or parenteral administration.

The term “aerosol” is used to denote various systems ranging from thoseof a colloidal nature to systems consisting of “pressurized packages.”The present definition refers to those products which depend upon thepower of a liquefied or compressed gas to dispense the activeingredient(s) in a finely dispersed mist, foam or semisolid. Pumpsystems which also dispense the active ingredient(s) in the form of afinally dispersed mist (although of greater particle size) often areclassified as aerosols.

The pressure package is convenient and easy to use. Medication isdispensed in a ready-to-use form at the push of a buttom. Since themedication is sealed in a tamper-proof pressure container, there is nodanger of contamination and the contents can be protected from air andmoisture. Easily decomposed drugs especially lend themselves to thistype of administration. For those products requiring regulation ofdosage, a metering valve can be used. An accurately measured dose oftherapeutically active drug can be administered quickly and in theproper particle-size range. In addition, when used with expensiveproducts, such as some steroids, savings can be achieved by the user ascompared to the use of other preparations such as ointments, creams orlotions. The aerosol dosage form allows for the dispensing of theproduct in the most desirable form; spray, foam or semisolid. Dependingon the nature of the product, the characteristics of the spray or foamcan be changed to insure the proper and most efficient use of themedication.

For parenteral application, particularly suitable vehicles consist ofsolutions, preferably oily or aqueous solutions, as well as suspensions,emulsions, or implants, including suppositories. Ampules are convenientunit dosages.

For enteral application, particularly suitable are tablets, dragees orcapsules having talc and/or a carbohydrate carrier binder or the like,the carrier preferably being lactose and/or corn starch and/or potatostarch. A syrup, elixir or the like can be used wherein a sweetenedvehicle is employed. Sustained release compositions can be formulatedincluding those wherein the active component is protected withdifferentially degradable coatings, e.g., by microencapsulation,multiple coatings, etc.

The nitrosated or nitrosylated compounds of the invention are used atdose ranges and over a course of dose regimen and are administered inthe same or substantially equivalent vehicles/carrier by the same orsubstantially equivalent oral or nasal inhalant devices as theirnon-nitrosated or non-nitrosylated counterparts. The nitrosated ornitrosylated compounds of the invention can also be used in lower dosesand in less extensive regimens of treatment. The amount of activeingredient that may be combined with the carrier materials to produce asingle dosage form will vary depending upon the host treated and theparticular mode of administration.

The dosage regimen for treating a disease condition with the compoundsand/or compositions of this invention is selected in accordance with avariety of factors, including the type, age, weight, sex, diet andmedical condition of the patient, the severity of the disease, the routeof administration, pharmacological considerations such as the activity,efficacy, pharmacokinetic and toxicology profiles of the particularcompound employed, whether a drug delivery system is utilized andwhether the compound is administered as part of a drug combination.Thus, the dosage regimen actually employed may vary widely and thereforemay deviate from the preferred dosage regimen set forth above.

Total daily dose administered to a host in single or divided doses maybe in amounts, for example, from about 1 to about 100 mg/kg body weightdaily and more usually about 3 to 30 mg/kg. Dosage unit compositions maycontain such amounts of submultiples thereof to make up the daily dose.

While the compounds of the invention can be administered as the soleactive pharmaceutical agent, they can also be used in combination withone or more compounds which are known to be effective against thespecific disease state targeted for treatment. The compositions of theinvention can also be administered as described above or can be made toinclude one or more additional active compounds which are known to beeffective against the specific disease state is targeted for treatment.

The invention also provides a pharmaceutical pack or kit comprising oneor more containers filled with one or more of the ingredients of thepharmaceutical compositions of the invention. Associated with suchcontainer(s) can be a notice in the form prescribed by a governmentalagency regulating the manufacture, use or sale of pharmaceuticals orbiological products, which notice reflects approval by the agency ofmanufacture, use or sale for human administration.

These and other aspects of the present invention will be apparent tothose skilled in the art from the teachings herein.

EXAMPLES Example 16α-Fluoro-11β-hydroxy-16α,17-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione-21-(3-methyl-3-nitrosothio)-butanoate1a. 3-Methyl-3(2,4,6-trimethoxyphenylmethylthio)butyric Acid

To a solution of 3-mercapto-3-methylbutyric acid (B. J. Sweetman et al.J. Med. Chem., 14, 868 (1971)) (4.6 g, 34 mmol) in methylene chloride(250 mL) under nitrogen and cooled over ice/salt to 5° C. (internaltemperature) was added trifluoroacetic acid (82 g, 0.72 mol). Nosignificant temperature rise was noted during the addition. To this wasthen added dropwise a solution of 2,4,6-trimethoxybenzyl alcohol (M. C.Munson et al., J. Org. Chem., 57, 3013 (1992)) (6.45 g, 32 mmol) inmethylene chloride (150 mL) such that the reaction temperature does notrise above 5° C. After the addition was complete, the mixture wasstirred for an additional 5 minutes at 5° C. and the volatiles wereremoved in vacuo (toluene or ethyl acetate can be used to assist in theremoval of volatile material). The residue was partitioned betweendiethyl ether and water and the organic phase dried over anhydroussodium sulfate, filtered and the volatile material removed in vacuo. Theresidue was treated with activated charcoal and recrystalised fromdiethyl ether/hexane. The product was isolated as an white solid in 70%yield (7 g) mp 103-105° C. ¹H NMR (CDCl₃) δ 6.12 (s, 2H), 3.80-3.85 (m,11H), 2.74 (s, 2H), 1.47 (s, 6H). ¹³C NMR (CDCl₃) δ173.9, 160.6, 158.6,105.6, 90.5, 55.7, 55.3, 45.9, 43.6, 28.4, 21.0.

1b.6α-Fluoro-11β-hydroxy-16α,17-((1-methylethylidene)bis(oxy))pregna-1,4-dien-3,20-dione-21-(3-methyl-3-(2,4,6-trimethoxyphenylmethylthio))-butanoate

A solution 6α-Fluoro-11β,21-dihydroxy-16α,17-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione (357 mg,0.8 mmol), the compound of Example 1a (251 mg, 0.8 mmol) and4-dimethylaminopyridine (20 mg, 0.16 mmol) in anhydrous DMF (5 mL) wastreated with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (157 mg, 0.8mmol). The resultant solution was stirred at room temperature for 2hours when the solvent was removed in vacuo and the residue poured intowater and extracted with ethyl acetate. The organic phase was dried overanhydrous sodium sulfate, filtered and the solvent removed in vacuo. Theresidue was chromatographed on silica gel using ethyl acetate:hexane(1:1) The product was isolated as a solid in 41% yield (240 mg), mp195-197° C. ¹H NMR (CDCl₃) δ 7.22 (d, J=10 Hz, 1H), 6.36 (s, 1H), 6.34(d, J=10 Hz, 1H), 6.14 (s, 2H), 5.35 (ddd, J=50 Hz, 11 Hz, 6 Hz),4.95-5.05 (m, 1H), 4.97 (d, J=18 Hz, 1H), 4.82 (d, J=18 Hz, 1H), 2H),4.47-4.55 (m, 1H), 3.81-3.89 (m, 11H), 2.86 (s, 2H), 2.46-2.54 (m, 1H),2.17-2.30 (m, 1H), 2.03-2.09 (m, 1H), 1.60-1.88 (m, 5H), 1.56 (s, 3H),1.54 (s, 3H), 1.45 (s, 6H), 1.1-1.5 (m, 5H), 1.24 (s, 3H), 0.96 (s, 3H).¹³C NMR (CDCl₃) δ 205.03, 186.82, 171.77, 166.52, 166.34, 161.65,160.03, 155.93, 129.67, 119.89, 119.72, 112.72, 108.78, 98.85, 92.03,89.87, 83.05, 71.02, 68.77, 57.12, 56.74, 56.11, 51.03, 47.80, 47.26,45.25, 45.02, 41.94, 41.31, 35.25, 30.01, 29.86, 29.46, 29.35, 27.95,27.05, 22.69, 22.21, 18.17. Analysis for C₃₉H₅₁FO₁₀S: 0.5H₂O:Calculated: C: 63.32; H, 7.03. Found: C: 63.39; H: 7.00.

1c.6α-Fluoro-11β-hydroxy-16α,17-((1-methylethylidene)bis(oxy))pregna-1,4-dien-3,20-dione-21-(3-methyl-3-mercapto)-butanoate

A stirred solution of the compound of Example 1b (220 mg, 27 μmol),anisole (200 μL, 1.8 mmol), phenol (200 mg, 2.1 mmol) and water (200 μL,11 mmol) in methylene chloride (0.2 mL) was treated with trifluoroaceticacid (1.5 mL). The resultant solution was stirred at room temperaturefor 1 hour. Toluene was added and the volatile material removed invacuo. The residue was chromatographed on silica gel using ethylacetate:hexane (1:3 then 1:1). The product was obtained as a solid in42% yield (70 mg) mp 205-208° C. ¹H NMR (CDCl₃) δ7.22 (d, J=9.8 Hz, 1H),6.36 (s, 1H) 6.34 (d, J=10 Hz, 1H), 5.35 (ddd, J=50 Hz, 11 Hz, 6 Hz),5.01 (d, J=4.2 Hz, 1H), 4.95 (d, J=6.2 Hz, 1H), 4.03-4.07 (m, 1H). 2.80(d, J=3 Hz, 2H), 2.46-2.54 (m, 1H), 2.43 (s, 1H), 2.17-2.30 (m, 1H),2.07-2.13 (m, 1H), 1.6-1.89 (m, 7H), 1.57 (s, 6H), 1.45 (s, 6H), 1.3-1.4(m, 2H), 1.24 (s, 3H), 1.16 (dd, J=1 Hz, 3.3 Hz, 1H), 0.96 (s, 3H).

1d.6α-Fluoro-11β-hydroxy-16α,17-((1-methylethylidene)bis(oxy))pregna-1,4-dien-3,20-dione-21-(3-methyl-3-nitrosothio)-butanoate

A solution of the compound of Example 1c (60 mg, 0.11 mmol) in methylenechloride (1 mL) was treated with tert butyl nitrite (25 μl, 22.5 mg,0.22 mmol) and the resultant solution stirred for 1 hr at roomtemperature. An additional 12 μL of tert butyl nitrite was added and thesolution stirred for an additional 15 min at room temperature. Thevolatile material was evaporated in vacuo and the residuechromatographed on silica gel using methanol:methylene chloride (1:25).The product was isolated as a green solid 40 mg (65%). ¹H NMR (CDCl₃) δ7.12 (d, J=10 Hz, 1H), 6.26 (s, 1H), 6.25 (d, J=10 Hz, 1H), 5.30 (ddd,J=50 Hz, 10 Hz, 6 Hz), 4.91-4.94 (m, 1H), 4.88 (d, J=18 Hz, 1H), 4.78(d, J=18 Hz, 1H), 4.41-4.47 (m, 1H), 3.33 (s, 2H), 2.38-2.47 (m, 1H),2.10-2.20 (m, 1H), 1.99 (s, 3H), 1.97 (s, 3H), 1.36 (s, 6H), 1.04-1.75(m, 8H), 1.15 (s, 3H), 0.86 (s, 3H).

Example 2 (8r)-3α-hydroxy-8-isopropyl-1αH,5αH-tropanium nitrite(±)-tropate

(8r)-3α-hydroxy-8-isopropyl-1αH,5αH-tropanium bromide (±)-tropate (0.2g, 0.48 mmol) was dissolved in hot water (2 mL) and to it was added asolution of silver nitrite (75 mg, 0.48 mmol) in boiling water (2 mL).The solution was stirred for 5 min. while keeping the solution hot andthen centrifuged at 1000 rpm for 5 min. The supernatant was decanted andthe water was removed by lyophilisation. The residue was recrystalisedfrom ethyl acetate/isopropanol to give the title compound as a whitesolid 124 mg (70%) mp 169-172° C. (dec). ¹H NMR (DMSO-d₆) δ 7.15-7.30(m, 5H), 4.90 (t, J=5.5 Hz, 1H), 3.97-4.04 (m, 1H), 3.65-3.82 (m, 4H),3.55-3.62 (m, 1H), 2.56 (s, 3H), 2.28-2.5 (m, 2H), 1.75-2.1 (m, 4H),1.57 (d, J=17 Hz, 1H), 1.40-1.50 (m, 1H), 1.06-1.14 (m, 6H). ¹³C NMR(DMSO-d₆) δ16.36, 24.46, 24.73, 31.25, 54.53, 55.18, 63.31, 65.20,65.30, 127.82, 128.47, 129.00, 136.48, 171.64. Anal calcd forC₂₀H₃₀N₂O₅:C: 63.47; H: 7.99; N, 7.40. Found: C: 63.28; H, 7.92; N,7.24.

Example 3 (8r)-3α-hydroxy-8-isopropyl-1αH,5αH-tropanium nitrate(O)-tropate

(8r)-3α-hydroxy-8-isopropyl-1αH,5αH-tropanium nitrate (O)-tropate (0.2g, 0.48 mmol) was dissolved in ethanol (2 mL) and to it added a solutionof silver nitrate (82 mg, 0.48 mmol) in water (2 mL). The solution wasstirred at room temperature for 1.5 hr. and centrifuged at 1000 rpm for5 min. The supernatant was decanted and the volatile materials removedby lyophilisation. The product was recrystalised from isopropanol togive the title compound as a white solid 150 mg (76%) mp 180-182° C. ¹HNMR (DMSO-d₆) δ 7.21-7.39 (m, 5H), 5.03 (t, J=5.7 Hz, 1H), 4.07-4.16 (m,1H), 3.78-3.94 (m, 4H), 3.59-3.65 (m, 1H), 2.68 (s, 3H), 2.4-2.6 (m,2H), 1.9-2.22 (m, 3H), 1.91 (d, J=17.2 Hz, 1H), 1.69 (d, J=17.2 Hz, 1H),1.49-1.59 (m, 1H), 1.06-1.14 (t, J=6 Hz, 6H). ¹³C NMR (DMSO-d₆) δ16.33,24.45, 24.73, 31.25, 54.52, 55.19, 63.32, 65.20, 65.31, 127.83, 128.48,129.00, 136.45, 171.62. Anal calcd for C₂₀H₃₀N₂O₆: C: 60.89; H: 7.66; N,7.10. Found: C: 60.71; H, 7.68; N, 6.90.

Example 49-Fluoro-11β-hydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione-21-(4-nitrooxy)-butanoate4a. 9-Fluoro-11β-hydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione-21-(4-bromo)-butanoateand9-Fluoro-11β-hydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione-21-(4-iodo)-butanoate

A solution of 9-Fluoro-11β, 21-dihydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione (1.6 g,3.7 mmol) and 4-bromobutyric acid (0.61 g, 3.7 mmol) in dry DMF (15 mL)at room temperature under nitrogen was treated with 4-DMAP (90 mg, 0.74mmol) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide methiodide (1.28g, 4.4 mmol) and the resultant solution stirred overnight at roomtemperature. The solvent was removed in vacuo and the residuechromatographed on silica gel using ethyl acetate:hexane (1:1 then 2:1)to give the product 0.8 g (an inseparable mixture of bromide and iodide)as a white solid together with 1.2 g of recovered 9-fluoro-11β,21-dihydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione.¹H NMR (CDCl₃) δ 7.23 (d, J=10 Hz, 1H), 6.37 (dd, J=10 Hz, 1.7 Hz, 1H),6.15 (s, 1H), 4.95 (dd, J=50 Hz, 18 Hz, 2H), 5.00 (s, 1H), 4.45 (br d,J=3.5 Hz), 3.53 (2t, J=6.4 and 6.7 Hz, 2H), 3.30 (2t, J=6.4 and 6.7 Hz,2H), 2.00-2.7 (m, 1H), 1.85-1.95 (m, 1H), 1.6-1.75 (m, 3H), 1.57 (s,3H), 1.45 (s, 3H), 1.24 (s, 3H), 0.96 (s, 3H).

4b. 9-Fluoro-11β-hydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione-21-(4-nitrooxy)-butanoate

To the products of Example 4a (0.45 g) in acetonitrile (5 mL) and DMF (1mL) was added silver nitrate (1.95 g 1.5 equivalents based on bromide asexclusive starting material) and the resultant solution stirredovernight at room temperature. Then, further silver nitrate (100 mg) wasadded and the resultant solution stirred an additional 24 hrs at roomtemperature. The solution was filtered and the filtercake washed withmethanol. The filtrate was reduced in vacuo and the residuechromatographed on silica gel using ethyl acetate:hexane (1:1) to givethe product as a white solid. mp. 211-213° C. ¹H NMR (CDCl₃) δ 7.26 (d,J=10 Hz, 1H), 6.35 (d, J=10 Hz, 1H), 6.14 (s, 1H), 4.95 (dd, J=50 Hz, 17Hz), 4.99 (d, J=4.6 Hz, 1H), 4.56 (t, J=6.2 Hz, 2H), 4.43 (d, J=8.5 Hz,1H), 2.33-2.75 (m, 7H), 2.04-2.18 (m, 3H), 1.84-1.95 (m, 1H), 1.55-1.75(m, 3H), 1.56 (s, 3H), 1.44 (s, 3H), 1.22 (s, 3H), 0.94 (s, 3H). Analcalcd for C₂₈H₃₆FNO₁₀: C, 59.46; H: 6.42. Found: C, 59.26; H, 6.34.

Example 59-Fluoro-11β-hydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione-21-(12-nitrosooxy)-dodecanoate5a. 9-Fluoro-11β-hydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene—3,20-dione-21-(12-tert-butyldimethylsilyloxy)-dodecanoate

A solution of 9-Fluoro-11β, 21-dihydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione (1 g, 2.3mmol) and 12-tert-butyldimethylsilyloxydodecanoic acid (760 mg, 2.3mmol) (J. Org. Chem., 50, 1616 (1985)) in dry DMF (10 mL) was stirred atroom temperature under nitrogen and 4-DMAP (56 mg, 0.46 mmol) addedfollowed by 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride(528 mg, 2.8 mmol). The resultant solution was stirred overnight at roomtemperature and the solvent removed in vacuo. The residue waschromatographed on silica gel using ethyl acetate:hexane (1:3 then 1:1)to give the product as a white solid 1.1 g (70% based on recoveredstarting material). mp. 146-148° C. ¹H NMR (CDCl₃) δ 7.16 (d, J=10 Hz,1H), 6.30 (d, J=10 Hz, 1H), 6.09 (s, 1H), 4.94 (d, J=4.9 Hz, 1H), 4.86(d, J=6.9 Hz), 4.37 (d, J=8.6 Hz, 1H), 3.55 (t, J=6.5 Hz, 2H), 2.59 (m,1H), 2.32-2.45 (m, 6H), 2.04-2.15 (m, 1H), 1.68-1.78 (m, 1H), 1.2-1.7(21H), 1.50 (s, 3H), 1.38 (s, 3H), 1.17 (s, 3H), 0.89 (s, 3H), 0.84 (s,9H), 0.00 (s, 6H).

5b. 9-Fluoro-11β-hydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione-21-(12-hydroxy)-dodecanoate

The product of Example 5a (0.15 g, 0.2 mmol) was dissolved in THF (1.3mL) and trifluoroacetic acid (160 μL) added followed by water (40 μL).The resultant solution was stirred at room temperature for 1 hr, thevolatile material removed in vacuo and the residue chromatographed onsilica gel using ethyl acetate:hexane (2:1) to give the product as awhite solid 100 mg (79%) mp 172-175° C. ¹H NMR (CDCl₃) δ7.21 (d, J=10Hz, 1H), 6.35 (dd, J=10 Hz, 1.7 Hz, 1H), 6.14 (s, 1H), 4.99 (d, J=4.7Hz, 1H), 4.91 (d, J=2.4 Hz, 2H), 4.41 (d, J=8.1 Hz, 1H), 3.65 (t, J=6.6Hz, 2H), 2.57-2.7 (m, 1H), 2.35-2.52 (m, 4H), 2.07-2.17 (m, 1H),1.83-1.94 (m, 1H), 1.44 (s, 3H), 1.57 (s, 3H), 1.55-1.77 (m, 7H),1.25-1.40 (m, 16H), 1.22 (s, 3H), 0.94 (s, 3H). ¹³C NMR (CDCl₃) δ203.82, 186.47, 173.69, 165.91, 152.04, 129.76, 125.13, 111.51, 101.24,98.91, 97.60, 81.88, 72.08, 71.57, 67.57, 62.92, 48.32, 48.02, 45.55,43.03, 36.91, 33.73, 33.68, 33.33, 33.08, 32.62, 30.88, 29.30, 29.21,29.18, 29.14, 28.99, 28.84, 27.55, 26.48, 25.80, 25.59, 24.75, 22.98,22.91, 16.29. Anal calcd for C₃₆H₅₃FO₈: C, 68.33; H, 8.44. Found: C,68.11; H, 8.37.

5c. 9-Fluoro-11β-hydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione-21-(12-nitrosooxy)-dodecanoate

The alcohol prepared above (90 mg, 142 μmol) was dissolved in thesolvent in a dry flask under N₂ and cooled over dry ice. After stirringfor 10 min pyridine (57.5 μL, 712 μmol) was added followed bynitrosonium tetrafluoroborate (33 mg, 285 μmol). The resultant solutionwas stirred at −78° C. for 3 hr, washed with water, dried over sodiumsulfate, filtered and the solvent removed in vacuo. The residue waschromatographed on silica gel using ethyl acetate:hexane:triethylamine(50:48:2) to give the product 45 mg (47%). ¹H. NMR (CDCl₃) δ 7.22 (d,J=10 Hz, 1H), 6.34 (dd, J=10 Hz, 1.3 Hz, 1H), 6.13 (m, 1H), 4.90 (dd,J=40 Hz, 18 Hz, 2H), 4.69 (br s, 2H), 4.41 (d, J=8 Hz, 1H), 2.28-2.71(m, 7H), 2.04-2.15 (m, 1H), 1.82-1.92 (m, 1H), 1.54 (s, 3H), 1.42 (s,3H), 1.21 (s, 3H), 1.2-1.8 (m, 22H), 0.94 (s, 3H). ¹³C NMR (CDCl₃) δ203.84, 186.43, 173.50, 165.81. 151.85, 129.86, 125.16, 111.51, 101.17,98.00, 81.86, 72.16, 71.66, 67.48, 48.26, 47.96, 45.48, 42.98, 37.27,33.77, 33.62, 33.27, 33.01, 30.86, 29.38, 29.32, 29.17, 29.12, 28.99,27.52, 26.46, 25.81, 25.76, 243.82, 22.97, 22.89, 16.36

Example 6 2-(tert-Butylamino-1-(4-hydroxy-3-hydroxymethylphenyl)ethanol2(R,S)-acetamido-3-methyl-3-nitrosothiol propionic acid salt 6a.2(R,S)-acetamido-3-methyl-3-nitrosothiol propionic acid

2(R,S)-acetamido-3-methyl-3-mercapto-propionic acid (0.2 g, 1.05 mmol)was dissolved in dry DMF (2 mL) and stirred at room temperature undernitrogen. tert-Butyl nitrite (248 μL, 215 mg, 2.09 mmol) was introducedand the resultant solution stirred at room temperature for 25 min whenthe solvent was removed in vacuo at room temperature. The green gum wasstirred with ether collecting the resultant solid by filtration (225 mg,97%). ¹H NMR (DMSO-d6) δ 13.21 (br s, 1H), 8.54 (d, J=9.45 Hz, 1H), 5.18(d, J=5.46 Hz, 1H), 1.98 (s, 3H), 1.95 (s, 3H), 1.88 (s, 3H).

6b. 2-(tert-Butylamino-1-(4-hydroxy-3-hydroxymethylphenyl)ethanol2(R,S)-acetamido-3-methyl-3-nitrosothiol propionic acid salt

2-(tert-Butylamino-1-(4-hydroxy-3-hydroxymethylphenyl)ethanol 25 mg, 0.1mmol) and the product of Example 6a (23 mg, 0.1 mmol) were stirred inwater to homogeneity then lyophilised to dryness to give the salt (48mg, 100%) mp 180-182° C. ¹H NMR (DMSO-d₆) δ 9.38 (br s, 1H), 7.81 (d,J=9.34 Hz, 1H), 7.33 (s, 1H), 7.08 (d, J=8.2 Hz, 1H), 6.75 (d, J=8.2 Hz,1H), 4.98 (br s, 1H), 4.69 (d, J=9.2 Hz, 1H), 2.72-2.95 (m, 2H), 1.97(s, 3H), 1.94 (s, 3H), 1.86 (s, 3H), 1.23 (s, 9H). ¹³C NMR (DMSO-d₆) δ172.52, 169.33, 154.07, 132.63, 128.60, 125.41, 125.29, 114.63, 69.16,61.45, 60.77, 58.61, 55.54, 48.91, 27.36, 26.21, 25.63, 23.04. Analcalcd for C₂₀H₃₃N₃O₇S: C, 52.27; H, 7.23; N, 9.14. Found: C, 52.49; H,7.33; N, 8.85.

Example 7 5,5′-((2-Hydroxy-1,3-propanediyl)bis-(oxy))bis(4-oxo-4H-1benzopyran-2(4-nitrooxybutyl)-carboxylic acid ester 7a.55′-((2-Hydroxy-1,3-propanediyl)bis-(oxy))bis(4-oxo-4H-1benzopyran-2(4-bromobutyl)-carboxylic acid ester

5,5′-((2-Hydroxy-1,3-propanediyl)bis-(oxy))bis(4-oxo-4H-1benzopyran-2-carboxylic acid disodium salt (2.6 g, 5.07 mmol) and1,4-dibromobutane (8.7 g, 40.6 mmol) were stirred at room temperature inDMSO for 72 hr and then the solvent removed by vacuum distillation. Theresidue was diluted with CH₂Cl₂ and washed with water. The aqueous phasewas extracted once with CH₂Cl₂ and the combined extracts washed withbrine and dried over sodium sulfate. The solution was filtered, thesolvent removed in vacuo and the residue chromatographed on silica gelusing ethyl acetate:hexane (4:1) to give cromolyn bis(4-ester) as asolid 1.2 g (32%). mp 130-133° C. ¹H NMR (DMSO-d₆) 67.74 (t, J=8.4 Hz,2H), 7.16 (dd, J=18.6 Hz, 8.4 Hz, 4H), 6.73 (s, 2H), 5.37 (br. s, 1H),4.29-4.40 (m, 9H), 3.63 (t, J=6.3 Hz, 4H), 1.83-2.03 (m, 8H). ¹³C NMR(DMSO-d₆) δ 176.88, 160.35, 158.77, 157.48, 150.40, 135.76, 115.73,114.96, 110.57, 109.38, 70.46, 67.34, 66.02, 35.08, 29.21, 26.93. Analcalcd for C₃₁H₃₀Br₂O₁₁: 50.42; H, 4.10. Found: C, 50.05; H, 4.42.

7b. 5,5′-((2-Hydroxy-1,3-propanediyl)bis-(oxy))bis(4-oxo-4H-1benzopyran-2(4-nitrooxybutyl)-carboxylic acid ester

The product of Example 7a (0.5 g, 0.68 mmol) was dissolved in a mixtureof acetonitrile (15 mL) and DMF (5 mL) and silver nitrate (0.92 g, 5.4mmol) added. The solution was stirred overnight at room temperature andthen additional silver nitrate (0.46 g, 2.7 mmol) added. After stirringan additional 24 hr at room temperature the solvent was removed in vacuoand the residue chromatographed directly on silica gel using ethylacetate:hexane (4:1) to give the product as a white solid. mp. 211-213°C. ¹H NMR (DMSO-d₆) δ 7.74 (t, J=8.4 Hz, 2H), 7.16 (dd, J=18.2 Hz, 8.4Hz, 2H), 6.73 (s, 2H), 5.37 (d, J=3.9 Hz), 4.57-4.63 (m, 4H), 4.28-4.40(m, 8H), 4.71 (t, J=6 Hz, 1H), 1.75-1.88 (m, 8H). ¹³C NMR (CDCl₃) δ178.03, 160.36, 158.75, 157.57, 150.25, 135.02, 116.32, 115.54, 111.05,109.63, 72.31, 70.25, 67.68, 65.81, 24.84, 23.55. Anal calcd forC₃₁H₃₀N₂O₁₇: 52.99; H, 4.30; N, 3.99. Found: C: 52.67; H, 4.38; N, 3.66.

Example 8 5,5′-((2-Hydroxy-1,3-propanediyl)bis-(oxy))bis(4-oxo-4H-1benzopyran-2(4-(2(S)-acetamido-3-methyl-3-nitrosothiol) propionic acidbutyl)-carboxylic acid ester 8a.5,5′-((2-Hydroxy-1,3-propanediyl)bis-(oxy))bis(4-oxo-4H-1benzopyran-2(4-(2(S)-tert-butyloxycarboxamido-3-methyl-3-tert-butyloxythiocarbonato)propionic acid butyl)-carboxylic acid ester

The product of Example 7a (0.7 g, 0.95 mmol) and2(S)-terbutyloxycaboxamido-3-methyl-3-tert-butyloxythiocarbonato-propionicacid (U.S. Pat. No. 5,187,305) (974 mg, 2.86 mmol) were mixed in dry DMF(10 mL) together with sodium iodide (28 mg, 0.19 mmol) and potassiumcarbonate (394 mg, 2.86 mmol). The resultant solution was stirred atroom temperature for 72 h, the solvent removed in vacuo and the residuechromatographed on silica gel using ethyl acetate:hexane (3:2) to givethe product as a foam 620 mg (52%). ¹H NMR (CDCl₆) δ 7.57 (t, J=8.4 Hz,2H), 7.14 (d, J=8.4 Hz, 2H), 6.94 (d, J=8.4 Hz, 2H), 6.92 (s, 2H), 5.58(d, J=9 Hz, 2H), 4.73 (d, J=7.4 Hz, 2H), 4.3-4.58 (m, 9H), 4.18-4.25 (m,4H), 1.77-1.92 (m, 8H), 1.50 (s, 6H), 1.45 (s, 18H), 1.44 (s, 6H), 1.40(s, 18H). ¹³C NMR (CDCl₃) δ 177.95, 170.52, 167.32, 160.32, 158.69,157.53, 155.16, 150.35, 134.86, 116.17, 115.51, 111.03, 109.54, 85.08,79.78, 70.22, 67.63, 66.08, 64.39, 59.88, 51.14, 28.17, 28.12, 27.80,25.32, 25.05, 25.01

8b.5,5′-((2-Hydroxy-1,3-propanediyl)bis-(oxy))bis(4-oxo-4H-1benzopyran-2(4-(2(S)-acetamido-3-methyl-3-mercapto)propionic acid butyl)-carboxylic acid ester

The product of Example 8a (0.62 mg, 0.49 mmol) was dissolved in CH₂Cl₂(9 mL) and anisole (450 mg) added followed by trifluoroacetic acid (10mL). The resultant solution was stirred at room temperature for 2 hrwhen the volatiles were removed in vacuo. The residue was dissolved inmore CH₂Cl₂ (9 mL) and stirred over ice under nitrogen. After 10 minutestriethylamine (286 μL, 206 mg, 2.04 mmol) was introduced followed byacetyl chloride (76 ?L, 84 mg, 1.07 mmol). The resultant solution wasstirred overnight at room temperature, washed with 0.5N HCl, dried overNa₂SO₄, filtered and the solvent removed in vacuo. The residue waschromatographed on silica gel using ethyl acetate:hexane (9:1) thenethyl acetate:methanol (9:1) to give the product as a foam 120 mg (30%).¹H NMR (CDCl₃) δ 7.54 (t, J=8.4 Hz, 2H), 7.08 (d, J=8.4, 2H), 6.90 (d,J=8.3 Hz, 2H), 6.87 (s, 2H), 6.57 (d, J=9.2 Hz, 2H), 4.58 (d, J=9.2 Hz,2H), 4.15-4.55 (m, 13H), 2.01 (s, 6H), 1.96 (s, 2H), 1.75-1.9 (s, 8H),1.44 (s, 6H), 1.31 (s, 6H). ¹³C NMR (CDCl₃) δ 177.88, 170.30, 169.91,160.18, 158.51, 157.37, 150.15, 134.84, 116.02, 115.25, 110.83, 109.29,70.09, 67.46, 65.93, 64.44, 60.33, 46.10, 30.83, 29.29, 24.90, 24.88,22.91.

8c. 5,5′-((2-Hydroxy-1,3-propanediyl)bis-(oxy))bis(4-oxo-4H-1benzopfran-2(4-(2(S)-acetamido-3-methyl-3-nitrosothiol) propionic acidbutyl)-carboxylic acid ester

The product of Example 8b (110 mg, 0.11 mmol) was dissolved in CH₂Cl₂ (3mL). To this was added tert butylnitrite (81 μL, 70 mg, 0.69 mmol) andthe resultant solution stirred at room temperature for 1 hr. Thevolatile material was removed in vacuo and the residue chromatographedon silica gel using ethyl acetate:acetone (4:1) to give the product as afoam 50 mg (43%). ¹H NMR (CDCl₃) δ 7.61 (t, J=8.4 Hz, 2H), 7.17 (d,J=8.4 Hz, 2H), 6.90-7.08 (m, 4H), 6.40 (d, J=9.2 Hz, 2H), 5.35 (d, J=9.3Hz, 2H), 4.1-4.65 (m, 13H), 2.07 (s, 6H), 2.04 (s, 6H), 1.95 (s, 6H),1.73-1.88 (m, 8H). ¹³C NMR (CDCl₃) δ 178.06, 169.95, 169.90, 160.38,158.76, 157.60, 150.35, 134.95, 116.27, 115.58, 111.09, 109.63, 70.28,67.70, 65.99, 65.04, 60.14, 57.87, 26.90, 26.54, 25.04, 24.93, 23.10.

Example 9 Comparative In Vivo Airway Responsiveness Activities

The measurement of biological activity in a pulmonary model of allergicasthma and lung inflammation was undertaken in adult conscious sheep.The methodology was essentially as described by W. M. Abraham et al.(Bull. Eur. Physiopathol. Respir., 22, 387 (1986)). Adult sheep with anatural cutaneous reaction to Ascaris Suum extract and who respond toinhalation challenge with Ascaris Suum with both an acutebronchoconstriction and a late bronchial obstruction were used in thestudy.

Unsedated sheep were intubated and measurement of airway mechanics werecomputed and recorded on a digital computer utilizing inputs oftranspulmonary pressure and tracheal airflow recordings. Each animal wasexposed to an aerosol of ascaris suum, diluted to produce an immediateand significant increase in mean lung resistance (sR1) of approximately600-700% above baseline. This acute response peaked within 30 minutesand remained significantly above baseline for 2 hours. By approximately6.5 hours after challenge sR1 again significantly increased overbaseline (130-200%) and remained elevated until 8 hours post challenge.This second response is known as the late response. sR1 returned tobaseline by 24 hours. Each animal was exposed to an aerosol of ascarissuum at a minimum of two week interval. Each animal was exposed to anaerosol of ascaris suum at a minimum of two week interval. Each animaldid not enter into a study until it responded in a reproducible mannerto three separate exposures of a designated dose of ascaris suum, eachexposure separated by a 2 week interval.

Airway responsiveness was measured by calculation of a PC150 value. ThePC150 is the dose of aerosolized carbachol which produces a 150%increase in sR1 above baseline. The doses of carbachol were administeredin an ascending dose-response fashion with each dose being administeredfor a 10 breath duration. The PC150 value was determined for each animal24 hours prior to exposure to ascaris suum and again 24 post exposure.The difference in PC150 values represented a change in airwayresponsiveness. The closer the post challenge to prechallenge ratio tounity the better the effect of the particular compound.

Each test drug was aerosolized to an animal 30 minutes prior tochallenge with ascaris suum.

Results

Sheep were treated with 500 μg of either Example 1 or 6α-flouro-11β,21-dihydroxy-16α,17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione. Data arereferenced to historical controls as described in the methods section.The data showed that Example 1 gave 59±14% protection of the immediateresponse and 89±2% protection of the peak late response (n=3).6α-Fluoro-11β, 21-dihydroxy-16α, 17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione gave 16±21% protection against theimmediate response and 83±7% protection against the peak late response(n=3).

Measurement of airway hyperresponsiveness showed the postchallenge/prechallenge ratios of PC150 values were 0.53±0.10 (control)and 1.18±0.06 (Example 1) and 0.59±0.06 (control) and 0.98±0.10(6α-Fluoro-11β, 21-dihydroxy-16α, 17α-((1-methylethylidene)bis(oxy))pregna-1,4-diene-3,20-dione).

The results show that Example 1 provides a better protection againstlung function changes due to aerosol challenge of ascaris suum in theconscious sheep.

It will be clear that the invention may be practiced otherwise than asparticularly described in the foregoing description and examples.Numerous modifications and variations of the present invention arepossible in light of the above teachings and, therefore, are within thescope of the appended claims.

1. A compound of Formula (III),

wherein, K is a a monovalent charged anion selected from halide,nitrate, or nitrite; R₁₂ and R₁₃ are hydrogen or, when taken togetherare oxygen; R₁₄ is lower alkyl or haloalkyl; R₁₅ is selected from:

wherein R_(v) is selected from D or M; D is (i) hydrogen (ii) —NO (iii)—NO₂ (iv) —CH(R_(d))—O—C(O)—Y—(C(R_(e))(R_(f)))_(p)-T-Q; (v)—C(O)-T¹-(C(R_(e))(R_(f)))_(p)-T²-Q; or (vi)—C(O)-T-(C(R_(y))(R_(z)))_(p); R_(d) is hydrogen, lower alkyl,cycloalkyl, aryl, alkylaryl or heteroaryl; Y is oxygen, sulfur orNR_(i); R_(i) is hydrogen, lower alkyl, lower haloalkyl or heteroaryl;R_(e) and R_(f) are each independently hydrogen, lower alkyl,cycloalkyl, aryl, heteroaryl, arylalkyl, alkylamino, dialkylamino, orcarboxy, or R_(e) and R_(f) taken together with the carbon atom to whichthey are attached are carbonyl, cycloalkyl or bridged cycloalkyl; p isan integer from 1 to 6; T, T¹ and T² are each independently a covalentbond, oxygen, sulfur or nitrogen; Q is —NO or —NO₂; with the provisothat in “T-Q” and “T²-Q”, T and T² cannot be a covalent bond; R_(y) andR_(z) are each independently-T¹-(C(R_(e))(R_(f)))_(p)-G-(C(R_(e))(R_(f)))_(p)-T²-Q; G is a covalentbond, -T-C(O)—, —C(O)-T or Y; wherein R_(d), R_(e), R_(f), p, Q, T, T¹,T² and Y are as defined herein; M is(i)—C(O)-T-(C(R_(e))(R_(f)))_(p)-G-(C(R_(e))(R_(f)))_(p)—N(N—(O—)N═O)—R₁or(ii)-CH(R_(d))—O—C(O)-T-(C(R_(e))(R_(f)))_(p)-G-(C(R_(e))(R_(f)))_(p)—N(N—(O—)N═O)—R₁;wherein T, R_(e), R_(f), p, G, R_(i), and R_(d) are as defined herein;and with the proviso that the compounds of Formula (III) must contain atleast one —NO group or at least one —NO₂ group, and wherein the at leastone —NO group or the at least one —NO₂ group is linked to the compoundsof Formula (II) through an oxygen atom, a nitrogen atom or a sulfuratom.
 2. The compound of claim 1, wherein the compound of Formula (III)is a nitrosated atropine, a nitrosylated atropine, a nitrosated andnitrosylated atropine, a nitrosated ipratropium, a nitrosylatedipratropium, a nitrosated and nitrosylated ipratropium, a nitrosatedflutropium, a nitrosylated flutropium, a nitrosated and nitrosylatedflutropium, a nitrosated tiotropium, a nitrosylated tiotropium or anitrosated and nitrosylated tiotropium.
 3. A composition comprising thecompound of claim 1 and a pharmaceutically acceptable carrier orexcipient.
 4. The composition of claim 3, wherein the composition is ina form that can be administered by oral inhalation, nasal inhalation,intranasal mucosal administration, orally, enterally, topically,vaginally, sublingually, rectally, intramuscularly, intravenously orsubcutaneously.
 5. The composition of claim 3, wherein the compositionis in form that can be administered as an aerosol.
 6. A method for (i)treating asthma; (ii) treating a respiratory disorder; or (iii) treatingcystic fibrosis in an individual in need thereof comprisingadministering an effective amount of the composition of claim
 3. 7. Themethod of claim 6, wherein the respiratory disorder is acute pulmonaryvasoconstriction, pneumonia, traumatic injury, aspiration or inhalationinjury, fat embolism in the lung, acidosis, inflammation of the lung,adult respiratory distress syndrome, acute pulmonary edema, acutemountain sickness, asthma, post cardiac surgery, acute pulmonaryhypertension, persistent pulmonary hypertension of the newborn, prenatalaspiration syndrome, hyaline membrane disease, acute pulmonarythromboembolism, heparin-protamine reactions, sepsis, statusasthmaticus, hypoxia, chronic pulmonary hypertension, bronchopulmonarydysplasia, chronic pulmonary thromboembolism, idiopathic pulmonaryhypertension, primary pulmonary hypertension or chronic hypoxia.
 8. Themethod of claim 6, comprising administering an effective amount of thecomposition by oral inhalation, by nasal inhalation, or by intranasalmucosal administration.
 9. The method of claim 6, comprisingadministering an effective amount of the composition as an aerosol. 10.A composition comprising a compound of Formula (III) and a compound thatdonates, transfers or releases nitric oxide, elevates endogenoussynthesis levels of nitric oxide or endothelium-derived relaxing factor,or is a substrate for nitric oxide synthase.
 11. The composition ofclaim 10, further comprising a pharmaceutically acceptable carrier orexcipient.
 12. The composition of claim 11, wherein the composition isin a form that can be administered by oral inhalation, nasal inhalation,intranasal mucosal administration, orally, enterally, topically,vaginally, sublingually, rectally, intramuscularly, intravenously orsubcutaneously.
 13. The composition of claim 11, wherein the compositionis in a form that can be administered as an aerosol.
 14. A method for(i) treating asthma; (ii) treating a respiratory disorder; or (iii)treating cystic fibrosis in an individual in need thereof comprisingadministering an effective amount of the composition of claim
 11. 15.The method of claim 14, wherein the respiratory disorder is acutepulmonary vasoconstriction, pneumonia, traumatic injury, aspiration orinhalation injury, fat embolism in the lung, acidosis, inflammation ofthe lung, adult respiratory distress syndrome, acute pulmonary edema,acute mountain sickness, asthma, post cardiac surgery, acute pulmonaryhypertension, persistent pulmonary hypertension of the newborn, prenatalaspiration syndrome, hyaline membrane disease, acute pulmonarythromboembolism, heparin-protamine reactions, sepsis, statusasthmaticus, hypoxia, chronic pulmonary hypertension, bronchopulmonarydysplasia, chronic pulmonary thromboembolism, idiopathic pulmonaryhypertension, primary pulmonary hypertension or chronic hypoxia.
 16. Themethod of claim 14, comprising administering an effective amount of thecomposition by oral inhalation, by nasal inhalation, or by intranasalmucosal administration.
 17. The method of claim 14, comprisingadministering an effective amount of the composition as an aerosol. 18.A kit comprising the composition of claim
 2. 19. A kit comprising acompound of Formula (III) and a compound that donates, transfers orreleases nitric oxide, elevates endogenous synthesis levels of nitricoxide or endothelium-derived relaxing factor, or is a substrate fornitric oxide synthase,
 20. The kit of claim 19, wherein the compound ofFormula (III) and the compound that donates, transfers or releasesnitric oxide, elevates endogenous synthesis levels of nitric oxide orendothelium-derived relaxing factor, or is a substrate for nitric oxidesynthase are separate components in the kit or are in the form of acomposition in the kit.